U.S. patent application number 15/724596 was filed with the patent office on 2018-02-01 for inhibitors of the kynurenine pathway.
This patent application is currently assigned to Curadev Pharma Pvt Ltd. The applicant listed for this patent is CURADEV PHARMA PRIVATE LTD.. Invention is credited to Monali Banerjee, Kamal Kishore Kapoor, Sandip Middya, Sushil Raina, Ritesh Shrivastava, Roger A. Smith, Arjun Surya, Scott K. Thompson, Aranapakam Venkatesan, Dharmendra B. Yadav, Veejendra K. Yadav.
Application Number | 20180030026 15/724596 |
Document ID | / |
Family ID | 51898766 |
Filed Date | 2018-02-01 |
United States Patent
Application |
20180030026 |
Kind Code |
A1 |
Banerjee; Monali ; et
al. |
February 1, 2018 |
INHIBITORS OF THE KYNURENINE PATHWAY
Abstract
The present application provides novel inhibitors of indoleamine
2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2 and/or
tryptophan 2,3-dioxygenase, metabolites thereof, and
pharmaceutically acceptable salts or prodrugs thereof. Also
provided are methods for preparing these compounds. A
therapeutically effective amount of one or more of the compounds of
formula (I) is useful in treating diseases resulting from
dysregulation of the kynurenine pathway. Compounds of formula (I)
act by inhibiting the enzymatic activity or expression of
indoleamine 2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2
and/or tryptophan 2,3-dioxy-genase.
Inventors: |
Banerjee; Monali; (Greater
Noida, IN) ; Middya; Sandip; (Greater Noida, IN)
; Shrivastava; Ritesh; (Greater Noida, IN) ;
Raina; Sushil; (Noida, IN) ; Surya; Arjun;
(New Delhi, IN) ; Yadav; Dharmendra B.; (Noida,
IN) ; Yadav; Veejendra K.; (Kanpur, IN) ;
Kapoor; Kamal Kishore; (Jammu, IN) ; Venkatesan;
Aranapakam; (Rego Park, NY) ; Smith; Roger A.;
(Chester Springs, PA) ; Thompson; Scott K.;
(Phoenixville, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CURADEV PHARMA PRIVATE LTD. |
Noida |
|
IN |
|
|
Assignee: |
Curadev Pharma Pvt Ltd
|
Family ID: |
51898766 |
Appl. No.: |
15/724596 |
Filed: |
October 4, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14774383 |
Sep 10, 2015 |
9815811 |
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PCT/US2014/024920 |
Mar 12, 2014 |
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15724596 |
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61782841 |
Mar 14, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 307/82 20130101; A61P 31/04 20180101; A61P 1/04 20180101; C07D
491/04 20130101; A61K 31/4436 20130101; A61P 11/04 20180101; C07D
409/04 20130101; A61K 31/444 20130101; A61P 25/20 20180101; Y02A
50/30 20180101; A61K 31/4545 20130101; A61P 25/28 20180101; A61K
31/343 20130101; A61K 31/4365 20130101; C07D 495/04 20130101; A61K
31/4355 20130101; A61P 1/16 20180101; A61P 31/18 20180101; A61K
31/704 20130101; G01N 2333/90241 20130101; A61P 1/18 20180101; A61P
15/02 20180101; Y02A 50/411 20180101; A61P 1/02 20180101; A61P
19/00 20180101; A61P 29/00 20180101; C07D 409/12 20130101; A61P
13/08 20180101; A61P 25/02 20180101; A61P 25/18 20180101; A61P
31/12 20180101; A61P 33/06 20180101; A61K 31/5377 20130101; A61P
37/00 20180101; A61P 13/10 20180101; A61P 35/02 20180101; A61K
31/381 20130101; A61P 17/00 20180101; A61P 37/04 20180101; C07D
491/048 20130101; C12Q 1/26 20130101; A61P 25/24 20180101; A61P
11/00 20180101; A61P 15/00 20180101; A61P 43/00 20180101; A61P
25/00 20180101; C07D 333/66 20130101; A61P 9/00 20180101; A61P
13/12 20180101; A61P 33/00 20180101; A61P 27/02 20180101 |
International
Class: |
C07D 333/66 20060101
C07D333/66; A61K 31/4355 20060101 A61K031/4355; A61K 31/4365
20060101 A61K031/4365; A61K 31/444 20060101 A61K031/444; A61K
31/4545 20060101 A61K031/4545; A61K 31/5377 20060101 A61K031/5377;
A61K 31/704 20060101 A61K031/704; A61K 31/381 20060101 A61K031/381;
C07D 409/12 20060101 C07D409/12; A61K 31/4436 20060101
A61K031/4436; C07D 491/048 20060101 C07D491/048; C12Q 1/26 20060101
C12Q001/26; C07D 307/82 20060101 C07D307/82; C07D 495/04 20060101
C07D495/04; C07D 491/04 20060101 C07D491/04; A61K 31/343 20060101
A61K031/343; C07D 409/04 20060101 C07D409/04 |
Claims
1-117. (canceled)
118. A compound of formula (1): ##STR00730## wherein: X.sup.1 is
CR.sup.1; X.sup.2 is CR.sup.2; X.sup.3 is N; X.sup.4 is CR.sup.4; Y
is O; Z is NR.sup.5R.sup.6; R', R.sup.2 and R.sup.4 are
independently selected from the group consisting of H, optionally
substituted C.sub.1-C.sub.6 alkyl, optionally substituted
C.sub.2-C.sub.6 alkenyl, optionally substituted C.sub.2-C.sub.6
alkynyl, optionally substituted C.sub.1-C.sub.6 alkoxy, mono or
bicyclic optionally substituted C.sub.6-C.sub.14 aryl, mono or
bicyclic optionally substituted heteroaryl, optionally substituted
tarypalkyl, (alkoxy)carbonyl, (alkyl)amido, (alkyl)amino,
optionally substituted mono or bicyclic cycloalkyl, optionally
substituted mono or bicyclic heterocyclyl, aminoalkyl,
alkylcarboxyl, (alkyl)carboxyamido, optionally substituted
(aryl)amino, hydroxyl, halogen, C.sub.1-C.sub.6 haloalkyl,
optionally substituted heterocyclyi(alkyl)-, optionally substituted
heteroaryk(alkyl), hydroxyalkyl, perfluoroalkyl, optionally
substituted aryloxy, optionally substituted heteroaryloxy,
optionally substituted C.sub.3-C.sub.8 cycloalkoxy,
N(R.sup.7).sub.2, CN, NO.sub.2, CO.sub.2H, CONR.sup.AR.sup.B,
S(O).sub.nR.sup.7, and optionally substituted heterocyclyloxy
having 1 to 2 heteroatoms selected from the group consisting of O,
S(O).sub.n, and NR.sup.7; n is 0 to 2; R.sup.A and R.sup.B are
independently selected from the group consisting of H, optionally
substituted C.sub.1-C.sub.6 alkyl, optionally substituted mono or
bicyclic C.sub.6-C.sub.14 aryl, optionally substituted mono or
bicyclic heteroaryl, optionally substituted (rypalkyl, optionally
substituted mono or bicyclic C.sub.3-C.sub.8 cycloalkyl, optionally
substituted mono or bicyclic heterocyclyl,
C.sub.1-C.sub.6haloalkyl, optionally substituted
heterocyclyl(alkyl), optionally substituted heteroaryl(alkyl),
hydroxyalkyl, and perfluomalkly; R.sup.5 is selected from the group
consisting of optionally substituted C.sub.1-C.sub.6 optionally
substituted mono or bicyclic C.sub.6-C.sub.14 aryl, optionally
substituted mono or bicyclic heteroaryl, optionally substituted
(aryl)alkyl, optionally substituted mono or bicyclic cycloalkyl,
optionally substituted mono or bicyclic heterocyclyl,
C.sub.1-C.sub.6 haloalkyl, optionally substituted
heterocyclyl(alkyl), optionally substituted heteroarykalkyl),
hydroxyalkyl, and perfluoroalkyl; R.sup.6 is H; and R.sup.7 is H,
C.sub.1-C.sub.6 alkyl, mono or bicyclic C.sub.6-C.sub.14 aryl, mono
or bicyclic heteroaryl, (arypalkyl, (alkoxy)carbonyl, (alkyl)amido,
(alkyl)amino, mono or bicyclic cycloalkyl, mono or bicyclic
heterocyclyl, alkylcarboxyl, heterocyclyl(alkyl),
heteroaryl(alkyl.), hydroxyalkyl, perfluoroalkyl, aryloxy,
heteroaryloxy C.sub.3-C.sub.6 cycloalkoxy, or heterocyclyloxy
having 1 to 2 heteroatoms selected from the group consisting of O,
S(O).sub.n, and NR.sup.A; or a pharmaceutically acceptable salt
thereof
119. The compound according to claim 118, wherein R.sup.1 is H,
halogen, CN, C.sub.1-C.sub.6 hydroxyalkyl, C.sub.1-C.sub.6 alkoxy,
or C.sub.1-C.sub.6 alkyl.
120. The compound according to claim 118, wherein R.sup.2 is
halogen, hydroxyl, CN, N(R.sup.7).sub.2, mono or bicyclic
optionally substituted C.sub.6-C.sub.14 aryl, optionally
substituted C.sub.1-C.sub.6 alkoxy, optionally substituted
C.sub.1-C.sub.6 alkyl., or optionally substituted aryloxy.
121. The compound according to claim 118, wherein R.sup.4 is H,
optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted C.sub.2-C.sub.6 alkenyl, optionally substituted
C.sub.2-C.sub.6 alkynyl, mono or bicyclic optionally substituted
C.sub.6-C.sub.14 aryl, mono or bicyclic optionally substituted
heteroaryl, optionally substituted (aryl)alkyl, optionally
substituted mono or bicyclic cycloalkyl, optionally substituted
mono or bicyclic heterocyclyl, halogen, optionally substituted
heterocyclyi(alkyl)-, optionally substituted heteroaryl(alkyl) or
CN.
122. The compound according to claim 121, wherein R.sup.4 is
optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted C.sub.2-C.sub.6 alkenyl, optionally substituted
C.sub.2-C.sub.6 alkynyl, optionally substituted (.sub.aryl)alkyl,
halogen, optionally substituted heterocyclyl(alkyl)-, optionally
substituted heteroaiyl(alkyl) or CN.
123. The compound according to claim 118, wherein R.sup.5 is
optionally substituted phenyl.
124. The compound according to claim 123, wherein R.sup.5 is:
##STR00731## wherein, R.sup.C to R.sup.G are independently selected
from the group consisting of H, halogen, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6alkoxy, heterocycle, optionally substituted
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, CN, --O(aryl),
C.sub.2-C.sub.6 alkynyl, C(O)C.sub.1-C.sub.6 alkyl,
--O--C.sub.3-C.sub.6 haloalkyl, and optionally substituted
aryl.
125. The compound according to claim 118, wherein R.sup.5 is
optionally substituted heteroaryl.
126. The compound according to claim 125, wherein R.sup.5 is
pyridine optionally substituted with one or more halogen.
127. The compound according to claim 118, wherein R is
benzo[d]dioxolane or tetrahydronaphthalene
128. The compound according to claim 118, selected from the group
consisting of:
N.sup.3-(3-Chloro-4-fluorophelayl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-Phenylfuro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-Chlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-(Trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,4-Dichlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Chloro-3-methoxyphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-Bromo-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Morpholinophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,4,5-Trifluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,4,5-Trifluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,4-Dimethylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,5-Dimethylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,4-Dichlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Bromophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Chlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-iodophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Ethylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Iodophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Bromo-4-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Bromo-4,5-difluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Benzoylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-(Piperidin-1-yl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(5-Chloro-2-methoxyphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-(Difluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Fluoro-3-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
3-((2-Aminofuro[2,3-c]pyridin-3-yl)amino)-2-fluorobenzonitrile,
N.sup.3-(4-Phenoxyphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-Chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,3,4-Trichlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,5-Di-tert-butylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,4-Dimethoxyphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(Benzo[d][1,3]dioxol-5-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Ethynylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,5-Dimethoxyphenyl)furo[2,3-c]pyridine-2,3-diamine,
4-((2-Aminofuro[2,3-c]pyridin-3-yl)amino)benzonitrile,
N.sup.3-(p-tolyl)Furo[2,3-c]pyridin-2,3-diamine,
1-(4-(t2-Aminofuro[2,3-c]pyridin-3-yl)amino)phenyl)ethanone,
N.sup.3-(3-Cyclopropylphenyl)furo[2,3-c]pridine-2,3-diamine,
N.sup.3-(3-Isopropylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-2,4-difluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,3-Dichloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Fluorophenylfuro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,5-Dichloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(m-tolyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(Pyridine-3-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-(Trifluoromethoxy)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(5-Chloro-2,4-difluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(5-Chloropyridine-3-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-([1,1'-Biphenyl]-3yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,4-Dimethylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2,5-Dichlorophenyl)furo[2,3-c]pyridine-2,3-diamine,
(2-Amino-3-((3-chloro-4-fluorophenyl)amino)-7-methylfuro[2,3-c]pyridine-4-
-methanol,
4-Chloro-N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Bromo-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Fluoro-3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diami-
ne, N.sup.3-(3-(tert-Butyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-Fluoro-3-(trifluoromethoxy)phenyl)furo[2,3-c]pyridine-2,3-diam-
ine,
N.sup.3-(5,6,7,8-Tetrahydronaphthalene-1-yl)furo[2,3-c]pyridine-2,3-d-
iamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-methoxyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-(Difluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-methylfuro[2,3-c]pyridine-2,3-diamine-
,
N.sup.3-(3-Chloro-4-fluorophenyl)-4-methoxyfuro[2,3c]pyridine-2,3-diamin-
e, N.sup.3-(3-Ethynylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-(1,1-Difluoroethyl)-4-fluorophenyl)furo[2,3-c]pyridine-2,3-dia-
mine,
N.sup.3-(3-Chloro-3-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(5,6,7,8-Tetrahydronaphthalene-2-yl)furo[2,3-c]pyridine-2,3-diami-
ne,
N.sup.3-(3-Chloro-4-fluorophenyl)-4,7-dimethylfuro[2,3-c]pyridine-2,3--
diamine,
N.sup.3(3-Chloro-4-fluorophenyl)-7-ethyl)furo[2,3-c]pyridine-2,3--
diamine,
7-Ethyl-N.sup.3-(4-fluoro-3-(trifluoromethyl)phenyl)furo[2,3-c]py-
ridine-2,3-diamine,
N.sup.3-(3-Benzylphenyl)furo[2,3-c]pyridine-2,3-diamine,
7-Ethyl-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diamin-
e,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-propylfuro[2,3-c]pyridine-2,3-diami-
ne,
7-Propyl-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-di-
amine,
N.sup.3-(4-Fluoro-3-(trifluoromethyl)phenyl)-7-propylfuro[2,3-c]pyr-
idine-2,3-diamine,
7-Methyl-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diami-
ne,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-phenylfuro[2,3-c]pyridine-2,3-diam-
ine,
7-Phenyl-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-d-
iamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-isopropylfuro[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-(Cyclopropylmethyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3(3-Chloro-4-fluorophenyl)-5-methoxy-7-methylfuro[2,3-c]pyridine-2,-
3-diamine,
7-Benzyl-N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Chloro-5-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Cyclohexylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-(Cyclobutylmethyl)phenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3-methoxyphenyl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(4-chlorophenyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Ethynylphenyl)-7-phenylfuro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-ethynylphenyl)-7-methylfuro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(pyridin-3-yl)furo[2,3-c]pyridine-2,3-
-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-chlorophenyl)furo[2,3-c]p-
yridine-2,3-diamine,
7-Ethyl-N.sup.3-(3-ethynylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine-2,3-
-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-methoxyphenyl)furo[2,3-c]-
pyridine-2,3-diamine,
3-((2-Aminofuro[2,3-c]pyridin-3-yl)amino)benzonitrile,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3-chlorophenyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(4-methoxyphenyl)furo[2,3-c]pyridine--
2,3-diamine,
5-Butoxy-N.sup.3-(3-chloro-4-fluorophenyl)-7-phenylfuro[2,3-c]pyridine-2,-
3-diamine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridine-7-
-carbonitile,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-phenoxy-7-phenylfuro[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-ethoxy-7-phenylfuro[2,3-c]pyridine-2,-
3-diamine,
5-(tert-Butoxy)-N.sup.3-(3-chloro-4-fluorophenyl)-7-phenylfuro[-
2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-isopropoxy-7-phenylfuro[2,3-c]pyridin-
e-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-cyclohexylfuro[2,3-c]pyridine-2,3-dia-
mine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-ethyl-7-phenylfuro[2,3-c]pyridin-
e-2,3-diamine,
N.sup.3-(3-Ethynyl-4-fluorophenyl)-7-phenylfuro[2,3-c]pyridine-2,3-diamin-
e,
N.sup.3-(3-Ethynyl-4-fluorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(pyridin-2-yl)furo[2,3-c]pyridine-2,3-
-diamine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)-7-(pyridine-2-yl)furo-
[2,3-c]pyridine-5-carbonitrile,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)-7-phenylfuro[2,3-c]pyridine-5--
carbonitrile,
N.sup.3-(3-Methoxy-5-methylphenyl)furo[2,3-c]pyridine-2,3-diamine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridine-7-carboxyli-
c acid,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridine-7-ca-
rboxamide,
N.sup.3-(3-Chloro-4-fluorophenyl)-N5,N5-dimethyl-7-phenylfuro[2-
,3-c]pyridine-2,3,5-triamine,
N.sup.3-(3-Cyclobutylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Cyclopentylphenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(trifluoromethyl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-phenoxyfuro[2,3-c]pyridine-2,3-diamin-
e,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(pyridine-2-yloxy)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-N.sup.7-phenylfuro[2,3-c]pyridine-2,3,7-
-triamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-N.sup.7-(pyridine-3-yl)furo[2-
,3-c]pyridine-2,3,7-triamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(phenylethenyl)furo[2,3-c]pyridine-2.-
3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-(2-(dimethylamino)ethoxy)-7-
-(pyridine-2-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-(3-dimethylamino)ethoxy)-7-phenylfuro-
[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(1-methyl-1H-pyrazol-5-yl)furo[2,3-c]-
pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(piperidin-1-yl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(4-methylpiperazin-1-yl)furo[2,3-c]py-
ridine-2,3-diamine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)-N-phenylfuro[2,3-c]pyridine-7--
carboxamide,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(4-fluorophenyl)furo[2,3-c]pyridine-2-
,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(1-methyl-1H-pyrazol-4-yl)furo[2,3-c]-
pyridine-2,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)benz-
onitrile,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin--
7-yl)benzamide, Methyl
4-(2-amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)benz-
oate,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-iodofuro[2,3-c]pyridine-2,3-diam-
ine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-morpholinopyridin-4-yl)furo[2
3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
7-Bromo-N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-d]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2,6-dimethylpyridin-4-yl)furo[2,3-c]-
pyridine-2,3-diamine,
7-Chloro-N3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-iodo-5-methoxyfuro[2,3-c]pyridine-2,3-
-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin--
7-yl)-N-(2-methoxyethyl)benzamide,
N.sup.3-(3-chloro-4-fluorophenyl)-7-(3,4-difluorophenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2,3,4-trifluorophenyl)furo[2,3-c]pyr-
idine-2,3-diamine,
7-Bromo-N.sup.3-(3-chloro-4-fluorophenyl)-5-methoxyfuro[2,3-c]pyridine-2,-
3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)-5-methoxyfuro[2,3-
-c]pyridin-7-yl)benzonitrile,
N.sup.3-(3-chloro-4-fluorophenyl)-7-(4-fluorophenyl)-5-methoxyfuro[2,3-c]-
pyridine-2,3-diamine, 4-(2-amino-3-((3-chloro-4-fluorophenyl)amino
-5-methoxyfuro[2,3-c]pyridin-7-yl)benzamide,
N.sup.3-(3-chloro-4-fluorophenyl)-7-fluorofuro[2,3-c]pyridine-2,3-diamine-
,
5-Chloro-N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamin-
e,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]p-
yridine-2,3-diamine,
N.sup.3-(3-chloro-4-fluorophenyl)-5-fluorofuro[2,3-c]pyridine-2,3-diamine-
,
2-Amino-3-((2-chlorophenyl)amino)furo[2,3-c]pyridine-7-carbonitrile,
2-Amino-3-((4-fluoro-3-(trifluoromethyl)phenyl)amino)furo[2,3-c]pyridine--
7-carbonitrile,
N.sup.3-(3,4-difluorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine-2,3-diam-
ine,
2-Amino-3-((3,4-difluorophenyl)amino)furo[2,3-c]pyridine-7-carbonitri-
le,
2-Amino-3-((3,5-difluorophenyl)amino)furo[2,3-c]pyridine-7-carbonitril-
e,
.sup.3-(3,5-Difluorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine-2,3-dia-
mine,
N.sup.3-(3,4-Difluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine,
7-Fluoro-N.sup.3-(4-fluoro-3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine-2,3-dia-
mine,
N.sup.3-(3-Chloro-4-fluorophenyl)7-(2,6-difluoropyridin-4-yl)furo[2,-
3-c]pyridine-2,3-diamine,
N.sup.3-(4-Fluoro-3-(trifluoromethyl)phenyl)-7-(pyridin-4-yl)furo[2,3-c]p-
yridine-2,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethylpicolinamide,
N.sup.3-(3,4-Difluorophenyl)-7-(2-methoxyphenyl)furo[2,3-c]pyridine-2,3-d-
iamine,
7-(2-Chlorophenyl)-N.sup.3-(3,4-difluorophenyl)furo[2,3-c]pyridine-
-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3-(trifluoromethyl)phenyl)furo[2,3-c-
]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-N7,N7-diphenylfuro[2,3-c]pyridine-2,3,7-
-triamine,
N.sup.3-(4-Fluorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine-2,-
3-diamine, N.sup.3-(2-Chlorophenyl)-7-
(pyridin-4-yl)furo[2,3-c]pyridine-2,3-diamine,
7-(Pyridin-4-yl)-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2-
,3-diamine, N.sup.3-(3-Chloro-4-fluorophenyl)-7-(naphthalen-
1-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-fluoro-3-(trifluoromethyl)phenyl)-7-(2-methylpyridin-4-yl)furo-
[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-chlorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3--
diamine,
7-(2-methylpyridin-4-yl)-N.sup.3-(3-(trifluoromethyl)phenyl)furo[-
2,3-c]pyridine-2,3-diamine,
N.sup.3-(4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,
3-c]pyridine-2,3-diamine,
N.sup.3-(3,4-difluorophenyl)-5-fluorofuro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chlorophenyl)-7-(pyridin-4-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(2-Chlorophenyl)-5-flurofuro[2,3-c]pyridine-2,3-diamine,
4-(2-Amino-3-((2-chlorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-methylpico-
linamide,
N.sup.3-(3-Chlorophenyl)-5-fluorofuro[2,3-c]pyridine-2,3-diamine- ,
5-Fluoro-N.sup.3-(3-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diam-
ine,
5-Fluoro-N.sup.3-(4-fluoro-3-(trifluoromethyl)phenyl)furo[2,3-c]pyrid-
ine-2,3-diamine,
4-(2-Amino-3-((3,4-difluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-methyl-
picolinamide,
4-(2-Amino-3-((4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-methylpico-
linamide,
4-(2-Amino-3-((3-chlorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethylpicolinamide,
4-(2-Amino-3-((3-(trifluoromethyl)phenyl)amino)furo[2,3-c]pyridin-7-yl)-N-
-methylpicolinamide,
4-(2-amino-3-((4-fluoro-3-(trifluoromethyl)phenyl)amino)furo[2,3-c]pyridi-
n-7-yl)-N-methylpicolinamide,
5-Fluoro-N.sup.3-(4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-methoxypyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine,
N.sup.3-(3-Chlorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3--
diamine,
7-(2-Methylpyridin-4-yl)-N3-phenylfuro[2,3-c]pyridine-2,3-diamine-
,
5-Fluoro-N.sup.3-(4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N.sup.3-(2-(piperidin-1-yl)phenyl)furo[2,3-c]pyr-
idine-2,3-diamine,
N.sup.3-(3-Fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3--
diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-fluoro-7-(2-methylpyridin-4-y-
l)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-morpholinofuro[2,3-c]pridine-2,3-diam-
ine,
N.sup.3-(3,4-Difluorophenyl)-5-fluoro-7-(2-methylpyridin-4-yl)furo[2,-
3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(naphthalen-2-yl)furo[2,3-c]pyridine--
2,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-p-
henylpropionamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-b-
utylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
tert-butyl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-i-
sobutylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-p-
ropylpicolthamide,
N.sup.3-(3-chlorophenyl)-5-fluoro-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
N.sup.3-(2,4-Difluorophenyl)-7-(2-methylpridin-4-yl)furo[2,3-c]pyridine-2-
,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-c-
yclohexylpicolinamide,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3,5-dichlorophenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
7-(2-(tert-Butyl)pyridin-4-yl)-N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3--
c]pyridine-2,3-diamine,
7-(Benzo[d][1,3]dioxol-5-yl)-N3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-ethylpyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(5-Chloro-2-fluorophenyl)-7-(2-methylpridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(3-Chloro-2-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-5-methyl-7-(2-methylpyridin-4-yl)furo[2-
,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-(trifluoromethyl)pyridin-4-yl)furo-
[2,3-c]pyridine-2,3-diamine,
(4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)pyr-
idin-2-yl)(piperidin-1-yl)methanone,
(4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)pyr-
idin-2-yl)(morpholino)methanone,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
pyridin-2-yl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
2-morpholinoethyl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-p-
entylpicolinamide,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2-chloro-5-fluorophenyl)furo[2,3-c]p-
yridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3,5-dimethylphenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(3,5-difluorophenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-styrylfuro[2,3-c]pyridine-2,3-diamine-
,
N.sup.3-(3-Bromo-4-methylphenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
N.sup.3-([1,1'-Biphenyl]-3-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridin-
e-2,3-diamine,
N.sup.3(3-Bromo-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridi-
n-2,3-diamine,
N.sup.3-(3-(1,1-Difluoroethyl)-4-fluorophenyl)-7-(2-methylpyridin-4-yl)fu-
ro[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-2,4-difluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]-
pyridine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N.sup.3-(4-(trifluoromethyl)phenyl)furo[2,3-c]py-
ridine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N.sup.3-(4-(trifluoromethoxy)phenyl)furo[2,3-c]p-
yridine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N.sup.3-(pyridin-3-yl)furo[2,3-c]pyridine-2,3-di-
amine,
N.sup.3-(4-Fluoro-3-(trifluoromethoxy)phenyl)-7-(2-methylpyridin-4--
yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3-(Difluoromethyl)phenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine,
3-((2-Amino-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridin-3-yl)amino)benzoni-
trile,
N.sup.3-(4-Chlorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridin-
e-2,3-diamine,
N.sup.3-Hexyl-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-Benzyl-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3-diamine,
N.sup.3-Cyclohexyl-7-(2-methylpyridin-4-yl)furo[2,3-c]pyridine-2,3-diamin-
e,
7-(2-Methylpyridin-4-yl)-N.sup.3-(tetrahydro-2H-pyran-4-yl)furo[2,3-c]p-
yridine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N.sup.3-((tetrahydro-2H-pyran-4-yl)methyl)furo[2-
,3-c]pyridine-2,3-diamine,
(E)-N.sup.3-(4-Fluorophenyl)-7-styrylfuro[2,3-c]pyridine-2,3-diamine,
(E)-N.sup.3-(3,4-Difluorophenyl)-7-styrylfuro[2,3-c]pyridine-2,3diamine,
(E)-N.sup.3-(4-Fluoro-3-(trifluoromethyl)phenyl)-7-styrylfuro[2,3-c]pyrid-
ine-2,3-diamine,
2-Amino-3-((3,4-difluorophenyl)amino)-N-methylfuro[2,3-c]pyridine-7-carbo-
xamide,
7-(Aminomethyl)-N.sup.3-(3-chlorophenyl)furo[2,3-c]pyridine-2,3-di-
amine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl
acetate,
N-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin--
7-yl)acetamide,
N.sup.3-(2-Chlorophenyl)-7-(piperidin-4-ylmethyl)furo[2,3-c]pyridine-2,3--
diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(pyridin-4-ylmethyl)furo[2,3-c]pyr-
idine-2,3-diamine,
(2-Amino-3-((3,4-difluorophenyl)amino)furo[2,3-c]pyridin-7-yl)methanol,
N.sup.3-(3-Chlorophenyl)-7-(trifluoromethyl)furo[2,3-c]pyridine-2,3-diami-
ne,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(cyclohexyloxy)furo[2,3-c]pyridine-
-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-((tetrahydro-2H-pyran-4-yl)oxy)furo[2,3-c]-
amine,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)-N-methylfuro[2,3-c]pyrid-
ine-7-sulfonamide,
2-Amino-3-((3,4-difluorophenyl)amino)-N-phenylfuro[2,3-c]pyridine-7-sulfo-
namide,
N.sup.3-(4-Fluorophenyl)-5-(trifluoromethyl)furo[2,3-c]pyridine-2,-
3-diamine,
N.sup.3-(2-Chlorophenyl)-5-(trifluoromethoxy)furo[2,3-c]pyridin-
e-2,3-diamine, 2-Amino
-3-((3-chloro-4-fluorophenyl)amino)-7-(2-methylpyridin-4-yl)furo[2,3-c]py-
ridine-5-carbonitrile,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N,N-
-dimethylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-e-
thylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-i-
sopropylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
2-methoxyethyl)picolinamide,
N-(3-(1H-Imidazole-1-yl)propyl)-4-(2-amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
pyridin-3-yl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
pyridin-4-yl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
4-methylpiperazin-1-yl)picolinamide,
2-Amino-3-((3-chloro-4-fluorophenyl)amino)-7-(2-methylpyridin-4-yl)furo[2-
,3-c]pyridin-5-ol,
N.sup.3-(3-chlorophenyl)-7-(2-fluoropyridin-4-yl)furo[2,3-c]pyridine-2,3--
diamine,
N.sup.3-(3-Fluorophenyl)-7-(2-fluoropyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(2-fluoropyridin-4-yl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(4-Fluorophenyl)-7-(2-fluoropyridin-4-yl)furo[2,3-c]pyridine-2,3--
diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(4-methylpridin-2-yl)furo[2,3-
-c]pyridine-2,3-diamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(imidazo[1,2-a]pyridin-5-yl)furo[2,3--
c]pyridiamine,
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(imidazo[1,2-c]pyridin-2-yl)furo[2,3--
c]pyridine-2,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethyl-1H-imidazole-1-carboxamide,
N.sup.3-(1-Methyl-1H-pyrrol-3-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
7-(2-Methylpyridin-4-yl)-N3-(1H-pyrrol-3-yl)furo[2,3-c]pyridine-2,3-diami-
ne,
N.sup.3-(1-Methyl-1H-pyrrol-2-yl-7-(2-methylpyridin-4-yl)furo[2,3-c]py-
ridine-2,3-diamine,
N.sup.3-(1-Methyl-1H-Indol-2-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(6-Chloropyrimidin-4-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(6-Fluoropyrimidin-4-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(2-Fluoropyrimidin-5-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
N.sup.3-(4,5-difluoropyrimidin-2-yl)-7-(2-methylpyridin-4-yl)furo[2,3-c]p-
yridine-2,3-diamine,
N.sup.3-(3-Chlorophenyl)-7-(2,6-difluoropyridin-4-yl)furo[2,3-c]pyridine--
2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(2,6-difluoropyridin-4-yl)furo[2,3-c]pyrid-
ine-2,3-diamine,
7-(2,6-Difluoropyridin-4-yl)-N3-(4-fluorophenyl)furo[2,3-c]pyridine-2,3-d-
iamine,
N.sup.3-(5-chloro-2-fluorophenyl)-7-(2,6-difluoropyridin-4-yl)furo-
[2,3-c]pyridine-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(4-(methoxymethyl)phenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
7-(2-Chloro-6-fluorophenyl)-N3-(3,4-difluorophenyl)furo[2,3-c]pyridine-2,-
3-diamine,
N-(4-(2-Amino-3-((4-difluorophenyl)amino)furo[2,3-c]pyridin-7-y-
l)phenyl)pyrrolidine-1 carboxamide,
N.sup.3-(3,4-Difluorophenyl)-7-(4-fluoro-2-methylphenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(3,4-Difluorophenyl)-7-(4-propoxyphenyl)furo[2,3-c]pyridine-2,3-d-
iamine,
N.sup.3-(3,4-Difluorophenyl)-7-(4-morpholinophenyl)furo[2,3-c]pyri-
dine-2,3-diamine,
5-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-2-f-
luorophenyl, Methyl
5-(2-amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-2-c-
hlorobenzoate,
3-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethylbenzamide,
4-(4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)p-
icolinamide)butanoic acid,
7-(2,6-Difluoropyridin-4-yl)-N.sup.3-(4-(trifluoromethyl)phenyl)furo[2,3--
c]pyridine-2,3-diamine,
7-(2-Fluoropyridin-4-yl)-N.sup.3-(4-(trifluoromethyl)phenyl)furo[2,3-c]py-
ridine-2,3-diamine,
5-Fluoro-7-(2-methylpyridin-4-yl)-N3-(4-(trifluoromethyl)phenyl)furo[2,3--
c]pyridine-2,3diamine,
4-(2-Amino-3-((4-(trifluoromethyl)phenyl)amino)furo[2,3-c]pyridin-7-yl)-N-
-methylpicolinamide,
4-(2-Amino-5-fluoro-3-((4-(trifluoromethyl)phenyl)amino)furo[2,3-c]pyridi-
n-7-yl)-N-methylpicolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)-5-fluorofuro[2,3-c]pyridin--
7-yl)-N-Methylpicolinamide,
5-Fluoro-N.sup.3-(4-(trifluoromethyl)phenyl)furo[2,3-c]pyridine-2,3-diami-
ne,
7-(2,6-Difluoropyridin-4-yl)-N.sup.3-(3-fluorophenyl)furo[2,3-c]pyridi-
ne-2,3-diamine,
N.sup.3-(2,5-Ddifluorophenyl)-7-(2,6-difluoropyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-(-
2-hydroxyethyl)picolinamide,
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N,N
diethyipicolinamide, and
N.sup.3-(3-Chloro-4-fluorophenyl)-7-(2,3-dichlorophenyl)furo[2,3-c]pyridi-
ne-2,3-diamine, or a pharmaceutically acceptable salt thereof.
129. A prodrug of a compound of claim 118, or a pharmaceutically
acceptable salt thereof, wherein the prodrug has a formula (II):
##STR00732## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, Y and Z
are as defined in claim 118, and R.sup.9 is optionally-substituted
C.sub.1-C.sub.6 alkyl or mono or bicyclic C.sub.6-C.sub.14 aryl or
heteroaryl.
130. A prodrug of a compound of claim 118, or a pharmaceutically
acceptable salt thereof, wherein the prodrug is a carbamate
compound which has a formula (III): ##STR00733## wherein X.sup.1,
X.sup.2, X.sup.3, X.sup.4, Y and Z are as defined in claim 118, and
R.sup.10 is optionally-substituted C.sub.1-C.sub.6 alkyl, aryl,
(aryl)alkyl, heteroaiyl, or (heteroarykalkyl.
131. The prodrug according to claim 130, which is: Benzyl
(3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carbamate;
Ethyl
(3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carbamat-
e, Methyl
(3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carba-
mate, Benzyl
(3-((3-chloro-4-fluorophenyl)amino)-7-cyanofuro[2,3-c]pyridin-2-yl)carbam-
ate, Ethyl
(7-(4-carbamoylphenyl)-3-((3-chloro-4-fluorophenyl)amino)furo[2-
,3-c]pyridin-2-yl)carbamate, Ethyl
(3-((3-chloro-4-fluorophenyl)amino)-7-phenylfuro[2,3-c]pyridin-2-yl)carba-
mate,or Ethyl
(3-((3-chloro-4-fluorophenyl)amino)-7-(2,6-dimethylpyridin-4yl)furo[2,3-c-
]pyridin-2-yl)carbamate, or a pharmaceutically acceptable salt
thereof.
132. A prodrug of a compound of claim 118, or a pharmaceutically
acceptable salt thereof, wherein the prodrug is a dicarbamate
compound which has a formula (IV): ##STR00734## wherein X.sup.1,
X.sup.2, X.sup.3, X.sup.4, Y and R.sup.5 are as defined in claim
118, and R.sup.11 is optionally-substituted C.sub.1-C.sub.6 alkyl,
aryl, (aryl)alkyl, heteroaryl, or (heteroarykalkyl.
133. The prodrug according to claim 132, which is: Benzyl
(2-(((benzyloxy)carbonyl)amino)furo[2,3-c]pyridin-3-yl)(3-chloro-4-fluoro-
phenyl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)furo[2,3-c]pyridin-3-y-
l)carbamate, Methyl
(3-chloro-4-fluorophenyl)(2-((methoxycarbonyl)amino)furo[2,3-c]pyridin-3--
yl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(7-cyano-2-((ethoxycarbonyl)amino)furo[2,3-c]pyr-
idin-3-yl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(7-(3-cyanophenyl)-2-((ethoxycarbonyl)amino)furo-
[2,3-c]pyridin-3-yl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-Iodofuro[2,3-c]pyri-
din-3-yl)carbamate, Ethyl
(7-((4-carbamoylphenyl)ethynyl)-2-((ethoxycarbonyl)amino)furo[2,3-c]pyrid-
in-3-yl)(3-chloro-4-fluorophenyl)carbamate, Ethyl
(7-bromo-2-((ethoxycarbonyl)amino)furo[2,3-e]pyridin-3-yl)(3-chloro-4-flu-
orophenyl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(7-(2,6-dimethylpridin-4-yl)-2-((ethoxycarbonyl)-
amino)furo[2,3-c]pyridin-3-yl)carbamate, Ethyl
(2-((ethoxycarbonyl)amino)-7-(pyridin-4-yl)furo[2,3-c]pyridin-3-yl)(4-flu-
oro-3-(trifluoromethyl)phenyl)carbamate, Ethyl
(7-cyano-2-((ethoxycarbonyl)amino)furo[2,3-c]pyridin-3-yl)(4-fluoro-3-(tr-
ifluoromethyl)phenyl)carbamate, Ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-(2-methylpyridin-4--
yl)furo[2,3-c]pyridin-3-yl)carbamate, or Ethyl
(3,4-difluorophenyl)(2-((ethoxycarbonyl)amino)-7-(2-methylpyridin-4-furo[-
2,3-c]pyridin-3-yl)carbamate, or a pharmaceutically acceptable salt
thereof.
134. A pharmaceutical composition, comprising a compound of claim
118, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier.
135. A method for regulating a kynurenine pathway, comprising the
step of administering a compound of claim 118, or a
pharmaceutically acceptable salt thereof, to a subject in need
thereof.
136. The method according to claim 135, wherein the method
comprises regulating one or more of indolamine 2,3-dioxygenase-1
(IDO1), indolamine 2,3-dioxygenase-2 (IDO2) and tryptophan
2,3-dioxygenase (TDO).
137. A method of treating cancer, bacterial infection, viral
infection, parasitic infection, immune-mediated disorder,
autoimmune disorder, inflammatory disease, central nervous system
disease, peripheral nervous system disease, neurodegenerative
disease, mood disorder, sleep disorder, cerebrovascular disease,
peripheral artery disease, cardiovascular disease, depression,
Alzheimer's disease, dementia, schizophrenia, HIV infection,
malaria, rheumatoid arthritis, insomnia or multiple sclerosis,
comprising the step of administering a compound of claim 118, or a
pharmaceutically acceptable salt thereof, to a subject in need
thereof
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This non-provisional application claims the benefit of U.S.
Provisional Application Ser. No. 61/782,841, filed on Mar. 14,
2013, which is incorporated by reference in its entirety.
FIELD OF INVENTION
[0002] The application generally relates to pharmaceutical
compounds that regulate enzymes indoleamine 2,3-dioxygenase-1
and/or indoleamine 2,3-dioxygenase-2 and/or tryptophan
2,3-dioxygenase and are useful for the treatment of diseases and
conditions with altered levels of the amino acid L-tryptophan and
metabolites of the kynurenine pathway.
BACKGROUND
[0003] The essential amino acid Tryptophan (Trp) is catabolized
through the kynurenine (KYN) pathway. The initial rate-limiting
step in the kynurenine pathway is performed by heme-containing
oxidoreductase enzymes, including tryptophan 2,3-dioxygenase (TDO),
indoleamine 2,3-dioxygenase 1 (IDO1), and indoleamine
2,3-dioxygenase 2 (IDO2). IDO1 and IDO2 share very limited homology
with TDO at the amino acid level and, despite having different
molecular structures, each enzyme has the same biochemical activity
in that they each catalyze tryptophan to form N-formylkynurenine.
IDO1, IDO2, and/or TDO activity alter local tryptophan
concentrations, and the build-up of kynurenine pathway metabolites
due to the activity of these enzymes can lead to numerous
conditions associated with immune suppression.
[0004] IDO1 and TDO are implicated in the maintenance of
immunosuppressive conditions associated with the persistence of
tumor resistance, chronic infection, HIV infection, malaria,
schizophrenia, depression as well as in the normal phenomenon of
increased immunological tolerance to prevent fetal rejection in
utero. Therapeutic agents that inhibit IDO1, IDO2, and TDO activity
can be used to modulate regulatory T cells and activate cytotoxic T
cells in immunosuppressive conditions associated with cancer and
viral infection (e.g. HIV-AIDS, HCV). The local immunosuppressive
properties of the kynurenine pathway and specifically IDO1 and TDO
have been implicated in cancer. A large proportion of primary
cancer cells have been shown to overexpress IDO1. In addition, TDO
has recently been implicated in human brain tumors.
[0005] The earliest experiments had proposed an anti-microbial role
for IDO1, and suggested that localized depletion of tryptophan by
IDO1 led to microbial death (Yoshida et al., Proc. Natl. Acad. Sci.
USA, 1978, 75(8):3998-4000). Subsequent research led to the
discovery of a more complex role for IDO1 in immune suppression,
best exemplified in the case of maternal tolerance towards the
allogeneic fetus where IDO1 plays an immunosuppressive role in
preventing fetal rejection from the uterus. Pregnant mice dosed
with a specific IDO1 inhibitor rapidly reject allogeneic fetuses
through induction of T cells (Munn et al., Science, 1998,
281(5380):1191-3). Studies since then have established IDO1 as a
regulator of certain disorders of the immune system and have
discovered that it plays a role in the ability of transplanted
tissues to survive in new hosts (Radu et al., Plast. Reconstr.
Surg., 2007 June, 119(7):2023-8). It is believed that increased
IDO1 activity resulting in elevated kynurenine pathway metabolites
causes peripheral and ultimately, systemic immune tolerance.
In-vitro studies suggest that the proliferation and function of
lymphocytes are exquisitely sensitive to kynurenines (Fallarino et
al., Cell Death and Differentiation, 2002, 9(10):1069-1077). The
expression of IDO1 by activated dendritic cells suppresses immune
response by mechanisms that include inducing cell cycle arrest in T
lymphocytes, down regulation of the T lymphocyte cell receptor
(TCR) and activation of regulatory T cells (T-regs) (Tcrness et
al., J. Exp. Med., 2002, 196(4):447-457; Fallarino et al., J.
Immunol., 2006, 176(11):6752-6761).
[0006] IDO1 is induced chronically by HIV infection and in turn
increases regulatory T cells leading to immunosuppression in
patients (Sci. Transl. Med., 2010; 2). It has been recently shown
that IDO1 inhibition can enhance the level of virus specific T
cells and concomitantly reduce the number of virus infected
macrophages in a mouse model of HIV (Potula et al., 2005, Blood,
106(7):2382-2390). IDO1 activity has also been implicated in other
parasitic infections. Elevated activity of IDO1 in mouse malaria
models has also been shown to be abolished by in vivo IDO1
inhibition (Tetsutani K., et al., Parasitology. 2007 7:923-30.
[0007] More recently, numerous reports published by a number of
different groups have focused on the ability of tumors to create a
tolerogenic environment suitable for survival, growth and
metastasis by activating IDO1 (Prendergast, Nature, 2011,
478(7368):192-4). Studies of tumor resistance have shown that cells
expressing IDO1 can increase the number of regulatory T cells and
suppress cytotoxic T cell responses thus allowing immune escape and
promoting tumor tolerance.
[0008] Kynurenine pathway and IDO1 are also believed to play a role
in maternal tolerance and immunosuppressive process to prevent
fetal rejection in utero (Munn et al., Science, 1998,
281(5380):1191-1193). Pregnant mice dosed with a specific IDO1
inhibitor rapidly reject allogeneic fetuses through suppression of
T cells activity (Munn et al., Science, 1998, 281(5380):1191-1193).
Studies since then have established IDO1 as a regulator of
immune-mediated disorders and suggest that it plays a role in the
ability of transplanted tissues to survive in new hosts (Radu et
al., Plast. Reconstr. Surg., 2007 June, 119(7):2023-8).
[0009] The local immunosuppressive properties of the kynurenine
pathway and specifically IDO1 and TDO have been implicated in
cancer. A large proportion of primary cancer cells overexpress IDO1
and/or TDO (Pilotte et al., Proc. Natl. Acad. Sci. USA, 2012, Vol.
109(7):2497-2502). Several studies have focused on the ability of
tumors to create a tolerogenic environment suitable for survival,
growth and metastasis by activating IDO1 (Prendergast, Nature,
2011, 478:192-4). Increase in the number of T-rcgs and suppression
of cytotoxic T cell responses associated with dysregulation of the
Kynurenine pathway by overexpression of IDO1 and/or TDO appears to
result in tumor resistance and promote tumor tolerance.
[0010] Data from both clinical and animal studies suggest that
inhibiting IDO1 and/or TDO activity could be beneficial for cancer
patients and may slow or prevent tumor metastases (Muller et al.,
Nature Medicine, 2005, 11(3):312-319; Brody et al., Cell Cycle,
2009, 8(12):1930-1934; Witkiewicz et al., Journal of the American
College of Surgeons, 2008, 206:849-854; Pilotte et al., Proc. Natl.
Acad. Sci. USA, 2012, Vol. 109(7):2497-2502). Genetic ablation of
the IDO1 gene in mice (IDO1-/-) resulted in decreased incidence of
DMBA-induced premalignant skin papillomas (Muller et al., PNAS,
2008, 105(44):17073-17078). Silencing of IDO1 expression by siRNA
or a pharmacological IDO1 inhibitor 1-methyl tryptophan enhanced
tumor-specific killing (Clin. Cancer Res., 2009, 15(2). In
addition, inhibiting IDO1 in tumor-bearing hosts improved the
outcome of conventional chemotherapy at reduced doses (Clin. Cancer
Res., 2009, 15(2)). Clinically, the pronounced expression of IDO1
found in several human tumor types has been correlated with
negative prognosis and poor survival rate (Zou, Nature Rev. Cancer,
2005, 5:263-274; Zamanakou et al., Immunol. Lett. 2007,
111(2):69-75). Serum from cancer patients has higher
kynurenine/tryptophan ratio, a higher number of circulating T-rcgs,
and increased effector T cell apoptosis when compared to serum from
healthy volunteers (Suzuki et al., Lung Cancer, 2010, 67:361-365).
Reversal of tumoral immune resistance by inhibition of tryptophan
2,3-dioxygenase has been studied by Pilotte et al. (Pilotte et al.,
Proc. Natl. Acad. Sci. USA, 2012, Vol. 109(7):2497-2502). Thus,
decreasing the rate of kynurenine production by inhibiting IDO1
and/or TDO may be beneficial to cancer patients.
[0011] IDO1 and IDO2 are implicated in inflammatory diseases. IDO1
knock-out mice don't manifest spontaneous disorders of classical
inflammation and existing kown small molecule inhibitors of IDO do
not elicit generalized inflammatory reactions (Prendergast et al.
Curr Med Chem. 2011; 18(15):2257-62). Rather, IDO impairment
alleviates disease severity in models of skin cancers promoted by
chronic inflammation, inflammation-associated arthritis and
allergic airway disease. Moreover, IDO2 is a critical mediator of
autoantibody production and inflammatory pathogenesis in autoimmune
arthritis. IDO2 knock-out mice have reduced joint inflammation
compared to wild-type mice due to decreased pathogenic
autoantibodies and Ab-secreting cells (Merlo et al. J. Immunol.
(2014) vol. 192(5) 2082-2090). Thus, inhibitors of IDO1 and IDO2
are useful in the treatment of arthritis and other inflammatory
diseases.
[0012] Kynurenine pathway dysregulation and IDO1 and TDO play an
important role in the brain tumors and are implicated in
inflammatory response in several neurodegenerative disorders
including multiple sclerosis, Parkinson's disease, Alzheimer's
disease, stroke, amyotrophic lateral schlerosis, dementia (Kim et
al., J. Clin. Invest, 2012, 122(8):2940-2954; Gold et al., J.
Neuroinflammation, 2011, 8:17; Parkinson's Disease, 2011, Volume
2011). Immunosuppression induced by IDO1 activity and the
Kynurenine metabolites in the brain may be treated with inhibitors
of IDO1 and/or TDO. For example, circulating T-reg levels were
found to be decreased in patient with glioblastoma treated with
anti-viral agent inhibitors of IDO1 (Soderlund, et al., J.
Neuroinflammation, 2010, 7:44).
[0013] Several studies have found Kynurenine pathway metabolites to
be neuroactive and neurotoxic. Neurotoxic kynurenine metabolites
are known to increase in the spinal cord of rats with experimental
allergic encephalomyelitis (Chiarugi et al., Neuroscience, 2001,
102(3):687-95). The neurotoxic effects of Kynurenine metabolities
is exacerbated by increased plasma glucose levels. Additionally,
changes in the relative or absolute concentrations of the
kynurenines have been found in several neurodegenerative disorders,
such as Alzheimer's disease, Huntington's disease and Parkinson's
disease, stroke and epilepsy (Nemeth et al., Central Nervous System
Agents in Medicinal Chemistry, 2007, 7:45-56; Wu et al. 2013; PLoS
One; 8(4)).
[0014] Neuropsychiatric diseases and mood disorders such as
depression and schizophrenia are also said to have IDO1 and
Kynurenine dysregulation. Tryptophan depletion and defficiency of
neurotransmitter 5-hydroxytryptamine (5-HT) leads to depression and
anxiety. Increased IDO1 activity decreases the synthesis of 5-HT by
reducing the amount of Tryptopan availability for 5-HT synthesis by
increasing Tryp catabolism via the kynurenine pathway (Plangar et
al. (2012) Neuropsychopharmacol Hung 2012; 14(4): 239-244).
Increased IDO1 activity and levels of both kynurenine and kynurenic
acid have been found in the brains of deceased schizophrenics
(Linderholm et al., Schizophrenia Bulletin (2012) 38: 426-432)).
Thus, inhibition of IDO1, IDO1, and TDO may also be an important
treatment strategy for patients with neurological or
neuropsychiatric disease or disorders such as depression and
schizophrenia as well as insomnia.
[0015] Kynurenine pathway dysregulation and IDO1 and/or TDO
activity also correlate with cardiovascular risk factors, and
kynurenines and IDO1 are markers for Atherosclerosis and other
cardiovascular heart diseases such as coronary artery disease
(Platten et al., Science, 2005, 310(5749):850-5, Wirlietner et al.
Eur J Clin Invest. 2003 Jul. 1; 33(7):550-4) in addition to kidney
disease. The kynurenines are associated with oxidative stress,
inflammation and the prevalence of cardiovascular disease in
patients with end-stage renal disease (Pawlak et al.,
Atherosclerosis, 2009, (204)1:309-314). Studies show that
kynurenine pathway metabolites are associated with endothelial
dysfunction markers in the patients with chronic kidney disease
(Pawlak et al., Advances in Medical Sciences, 2010,
55(2):196-203).
[0016] There is a need in the art for compounds that are inhibitors
of the indoleamine 2,3-dioxygenase-1 and/or indoleamine
2,3-dioxygenase-2 and/or tryptophan 2,3-dioxygenase pathway, as
well as for methods for treating diseases that can benefit from
such inhibition.
SUMMARY OF THE INVENTION
[0017] In one aspect, the present invention provides a compound of
formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof, wherein X.sup.1-X.sup.4, Y, and
Z are defined herein.
##STR00001##
[0018] In another aspect, compounds of formulae (I-A), (I-AA),
(I-AAA), (I-B), (I-BB), (I-BBB), (I-BBBB), (I-BBBBB), (I-BBBBBB),
(I-C), (I-CC), (I-D), (I-DD), (I-E), (I-EE), (I-F), (I-FF), (I-C)
and (I-CC) are provided, wherein X.sup.1-X.sup.4, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are defined herein.
##STR00002## ##STR00003## ##STR00004##
[0019] In another aspect, the invention relates to prodrugs of
compounds of formula (I) having a formula (II), (III) or (IV) are
provided, wherein R.sup.9-R.sup.11 are defined herein
##STR00005##
[0020] In one aspect, the invention relates to a metabolite of a
compound of formula (I) (IV) or a prodrug of said compound of
formula (I), or a pharmaceutically acceptable salt or a prodrug
thereof.
[0021] In a further aspect, a composition comprising a compound or
prodrug thereof as described herein and a pharmaceutically
acceptable carrier is provided.
[0022] In another aspect, a composition comprising a metabolite of
a compound of formula (I) (IV) or prodrug thereof as described
herein and a pharmaceutically acceptable carrier is provided,
[0023] In yet another aspect, a kit comprising a compound of
formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof described herein is
provided.
[0024] In another aspect, a method for treating a disease treatable
by inhibiting a kynurenine pathway is provided and includes
administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to a subject in
need thereof. In another aspect, a method for regulating a
kynurenine pathway is provided and includes administering a
compound or a metabolite thereof, or a pharmaceutically acceptable
salt or prodrug thereof as described herein to a subject in need
thereof.
[0025] In another aspect, a method of regulating one or more of a
indoleamine 2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2
or a tryptophan 2,3-dioxygenase enzymes is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or prodrug thereof as described
herein to a subject in need thereof. In another aspect, the
regulating is inhibiting the kynurenine pathway or one or more of
the enzymes.
[0026] In still a further aspect, a method of regulating the
kynurenine pathway by inhibiting indoleamine 2,3-dioxygenase-1
and/or indoleamine 2,3-dioxygenase-2 and/or tryptophan
2,3-dioxygenase is provided and includes administering a compound
or a metabolite thereof, or or a pharmaceutically acceptable salt
or prodrug thereof as described herein to a subject in need
thereof
[0027] In one aspect, a method of reducing kynurenine pathway
metabolites is provided and includes administering a compound or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof.
[0028] In another aspect, a method of altering tryptophan levels in
a subject and includes administering a compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof described herein is provided. In one aspect, the
tryptophan levels are increased. In another aspect,
kynurenine/tryptophan ratio is decreased.
[0029] In one aspect, a method of treating a disease associated
with or resulting from dysregulation of a kynureninc pathway is
provided and includes administering a compound of formula (1) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof.
[0030] In another aspect, a method for treating a disease caused by
the dysregulation of the kynurenine pathway by inhibiting
indoleamine 2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2
and/or tryptophan 2,3-dioxygenase is provided and includes
administering a compound or a pharmaceutically acceptable salt or
prodrug thereof described herein to a subject in need thereof.
[0031] In yet another aspect, a method for treating a disease
caused by activation of indoleamine 2,3-dioxygenase-1 or tryptophan
2,3-dioxygenase or both enzymes is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or prodrug thereof as described
herein to a subject in need thereof. In yet another aspect, a
method for treating a disease caused by activation of indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes or both is
provided and includes administering a compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof.
[0032] In yet another aspect, a method for treating a disease
caused by activation of indolcaminc 2,3-dioxygenase-1 or
indoleamine 2,3-dioxygenase-2 or both is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or prodrug thereof as described
herein to a subject in need thereof.
[0033] In another aspect, a method of inhbiting activation of one
or more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes is provided
and includes administering a compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof
[0034] In another aspect, a method for treating a disease
associated with one or more of indoleamine 2,3-dioxygenase-1 or
indoleamine 2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes
is provided and includes administering a compound of formula (I) or
a metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof described herein to a subject in need thereof.
[0035] In yet a further aspect, a method is provided for treating a
disease characterized by abnormal immune suppression resulting from
dysregulation of a kynurenine pathway and includes administering a
compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a subject in need thereof.
[0036] In a further aspect, a method for regulating a disease
characterized by abnormal immune suppression resulting from a
dysregulated kynurenine due to activation any one or more of
indoleamine 2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or
tryptophan 2,3-dioxygenase enzyme is provided and includes
administering a compound, a metabolite thereof or a
pharmaceutically acceptable salt or prodrug thereof as described
herein to a subject in need thereof.
[0037] In one aspect, a method of treating immune suppression is
provided and includes administering a compound or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
as described herein to a subject in need thereof In another aspect,
the immune suppression is associated with increased kynurenine
metabolite levels or enzymatic activity of one or more of
indoleamine 2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or
tryptophan 2,3-dioxygenase enzymes. In yet another aspect, a method
is provided for treating immune suppression associated with one or
more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes and
includes administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to a subject in
need thereof Thus, in an aspect, compounds of the invention for use
in treatment of immunosuppression associated with one or more of
indoleamine 2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or
tryptophan 2,3-dioxygenase enzymes are provided.
[0038] In yet another aspect, a method for treating immune
suppression through inhibiting enzymatic activity of indoleamine
2,3-dioxygenase and/or tryptophan 2,3-dioxygenase is provided and
includes administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a subject in need thereof
[0039] In yet another aspect, a method of reducing or eliminating
an immune mediated disorder is provided and includes administering
a compound of formula (1) (IV) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a patient.
[0040] In yet another aspect, a method of inhibiting an autoimmune
reaction or autoimmune antibody production in a subject is provided
and includes administering a compound or a metabolite thereof; or a
pharmcetucially acceptable salt or prodrug thereof as described to
a subject in need thereof. In another aspect, a method of
inhibiting autoimmune reaction or autoimmune antibody production is
inhibited by (i) inhibiting indoleamine 2,3-dioxygenase-2 or (ii)
reducing kynurenine metabolites, and includes administering a
compound or a metabolite thereof, or a pharmcetucially acceptable
salt or prodrug thereof as described to a subject in need thereof.
In one aspect, the foregoing reduction in autoimmune reaction or
autoimmune antibody production is associated with inflammatory
diseases, cancer or autoimminc disorders.
[0041] In one aspect, the list of diseases comprise cancer,
bacterial infection, viral infection, parasitic infection,
immune-mediated disorder, autoimmune disorder, inflammatory
disease, central nervous system disease, peripheral nervous system
disease, neurodegenerative disease, mood disorder, sleep disorder,
cerebrovascular disease, peripheral artery disease, or
cardiovascular disease. In another aspect, all foregoing methods
comprise administration of one or more therapeutic agent or
therapy. In one aspect, the therapeutic agent is a chemotherapeutic
agent selected from a group further comprising a cancer vaccine, a
targeted drug, a targeted antibody, an antibody fragment, an
antimetabolite, an antineoplastic, an antifolate, a toxin, an
alkylating agent, a DNA strand breaking agent, a DNA minor groove
binding agent, a pyrimidine analog, a purine analog, a
ribonucleotide reductase inhibitor, a tubulin interactive agent, an
anti-hormonal agent, an immunomoldulator, an anti-adrenal agent, a
cytokine, a radiation therapy, a cell therapy, or a hormone
therapy.
[0042] In another aspect, a method of treating depression,
Alzheimer's disease, dementia, schizophrenia, HIV inection,
malaria, rheumatoid arthritis, insomnia or multiple sclierosis is
provided and include administering a compound or a metabolite
thereof; or a pharmaceutically acceptable salt or prodrug thereof
described herein to a patient.
[0043] In one aspect, the disease is cancer. In another aspect,
cancer disease is a cancer of squamous cell, peritoneum, prostate,
head, neck, eye, mouth, throat, esophagus, bronchus, larynx,
pharynx, thyroid cancer, chest, bone, lungs, colon, rectum,
stomach, urinary bladder, gall bladder, uterus, cervix, breast,
ovaries, uterus, vagina, vulva, testicles, penis, anus, skin,
thyroid, blood, lymph nodes, kidney, liver, intestines, salivary
gland, pancreas, brain, spine, adrenal gland, skin or leukemia. In
another aspect, a method of treating tumor resistance is provided
comprising administering a compound or a metabolite thereof; or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a patient.
[0044] In another aspect, the viral infection is HIV infection. In
another aspect, parasite infection is malaria or Leishmaniasis.
[0045] In another aspect, a method of preparing a compound of
formula (I-C) is provided as described herein. In another aspect,
compounds obtainable by the method of preparing a compound of
formula (I-C) are provided.
[0046] In yet another aspect, a method for diagnosing and treating
a disease associated with kynurenine pathway or one or more of
indoleamine 2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2
or a tryptophan 2,3-dioxygenase enzymes in a subject is provided
and includes: (i) assaying a blood and/or tissue sample from a
subject; (ii) determining the subject's blood and/or tissue
tryptophan or Kynurenine concentration or both in the sample; (iii)
optionally determining the subject's Kynurnine/tryptophan ratio;
and (iv) administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a subject.
[0047] In still another aspect, a method of monitoring a disease
associated with kynurenine pathway or one or more of indoleamine
2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2 or a
tryptophan 2,3-dioxygenase enzymes in a subject is provided and
includes (i) dosing a subject having a disease associated with
kynurenine pathway with a compound, (ii) analyzing a blood or
tissue samples or both at one or more time points or continuously
during a treatment regimen, (iii) determining a tryptophan and a
kynurenine concentration in the blood or the tissue sample or both,
(iv) optionally determining the subject's kynumine/tryptophan
ratio, and (v) adjusting the treatment regimen or dosage of the
compound.
[0048] In a further aspect, a method for diagnosing and treating a
disease associated with kynurenine pathway or one or more of
indoleamine 2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2
or a tryptophan 2,3-dioxygenase enzymes in a patient is provided
and includes (i) analyzing a patient sample for the presence or
absence of altered kynurenin/tryptophan ratio, wherein the patient
is diagnosed with a disease associated with kynurenine pathway if
altered kynurenine/tryptophan ratio is detected and (ii)
administering a compound to the diagnosed patient.
[0049] In still a further aspect, a method for treating a disease
associated with kynurenine pathway or one or more of an indoleamine
2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2 or a
tryptophan 2,3-dioxygenase enzyme in a patient and includes (i)
requesting a test providing the results of an analysis to determine
whether the patient's kynurnine levels are altered, and (ii)
administering a compound to the patient if the patient's kynurenine
levels are altered.
[0050] In yet another aspect, a use of foregoing methods is
provided wherein the disease is cancer, bacterial infection, viral
infection, parasitic infection, immune-mediated disorder,
autoimmune disorder, inflammatory disease, central nervous system
disease, peripheral nervous system disease, neurodegenerative
disease, mood disorder, sleep disorder, cerebrovascular disease,
peripheral artery disease, or cardiovascular disease.
[0051] Other aspects and advantages of the invention will be
readily apparent from the following detailed description of the
invention.
BRIEF DESCRIPTION OF DRAWINGS
[0052] FIG. 1(a). Shows the mean tumor growth rate of CT-26 tumor
cells in Balb/c mice when dosed orally BID with either test
compound 2 (40 mg/kg in 30% PEG400+20% PG in NS) or vehicle
alone.
[0053] FIG. 1(b). Shows the mean tumor growth rate of CT-26 tumor
cells in Balb/c mice when dosed orally BID with either test
compound 97 (75 mg/kg in 40% PEG400+20% PG+10% DACM in NS) or
vehicle alone.
[0054] FIG. 1(c). Shows the mean tumor growth rate of CT-26 tumor
cells in Balb/c mice when dosed orally BID with either test
compound 166 (60 mg/kg in 40% PEG400+20% PG+10% DACM in NS) or
vehicle alone.
[0055] FIG. 1(d). Shows the mean tumor growth rate of CT-26 tumor
cells in Balb/c mice when dosed orally BID with either test
compound 184 (50 mg/kg in 40% PEG400+20% PG+10% DACM in NS) or
vehicle alone.
[0056] FIG. 2(a). Shows the mean tumor growth rate of CT-26 cells
in Balb/c mice when dosed orally BID with test compound 184 (50
mg/kg in 40% PEG400+20% PG+10% DACM in NS) and vehicle either alone
or in combination with Doxorubicin (DOXO).
[0057] FIG. 2(b). Shows the mean tumor growth rate of CT-26 tumor
cells in Balb/c mice when dosed orally BID with test compound 97
(75 mg/kg in 40% PEG400+20% PG+10% DACM in NS) and vehicle either
alone or in combination with DOXO.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0058] The following definitions are used in connection with the
compounds of the present invention unless the context indicates
otherwise.
[0059] Throughout the description and the claims of this
specification the word "comprise" and other forms of the word, such
as "comprising" and "comprises," means including but not limited
to, and is not intended to exclude for example, other additives,
components, integers, or steps.
[0060] As used in the description and the appended claims, the
singular forms "a," "an," and "the" include plural referents unless
the context clearly dictates otherwise. Thus, for example,
reference to "a composition" includes mixtures of two or more such
compositions.
[0061] "Optional" or "optionally" means that the subsequently
described event or circumstances can or cannot occur, and that the
description includes instances where the event or circumstance
occurs and instances where it does not.
[0062] The following definitions are used in connection with the
compounds of the present invention unless the context indicates
otherwise. In general, the number of carbon atoms present in a
given group is designated "C.sub.x-C.sub.y", where x and y arc the
lower and upper limits, respectively. For example, a group
designated as "C.sub.1-C.sub.6" contains from 1 to 6 carbon atoms.
The carbon number as used in the definitions herein refers to
carbon backbone and carbon branching, but does not include carbon
atoms of the substituents, such as alkoxy substitutions and the
like. Unless indicated otherwise, the nomenclature of substituents
that are not explicitly defined herein are arrived at by naming
from left to right the terminal portion of the functionality
followed by the adjacent functionality toward the point of
attachment. For example, the substituent "arylalkyloxycarbonyl"
refers to the group (C.sub.6-C.sub.14 aryl)-(C.sub.1-C.sub.6
alkyl)-O--C(O)--. The term optionally substituted refers to
replacing a hydrogen atom of a group with an alkyl, alkoxy, aryl,
monocyclic or bicyclic cycloalkyl, mono or bicyclic
heterocyclylalkyl, (aryl)alkyl, (alkoxy)carbonyl, (alkyl)amido,
(alkyl)amino, --NH.sub.2, aminoalkyl, alkylcarboxyl,
(alkyl)carboxyamido, (aryl)amino, haloalkyl, heteroaryl,
heterocyclyl, heteroaryl(alkyl), mono, di or perfluoroalkyl,
halogen, CN, C(O)OH, amide, amide formed from a primary or
secondary amine, NO.sub.2, OH, mono-fluoroalkoxy, di-fluoroalkoxy,
trifluoroalkoxy, and hydroxyalkyl. Terms not defined herein have
the meaning commonly attributed to them by those skilled in the
art.
[0063] "Alkyl" refers to a hydrocarbon chain that may be a straight
chain or branched chain, containing the indicated number of carbon
atoms, for example, a C.sub.1-C.sub.12 alkyl group may have from 1
to 12 (inclusive) carbon atoms in it. Examples of C.sub.1-C.sub.6
alkyl groups include, but are not limited to, methyl, ethyl,
propyl, butyl, pentyl, hexyl, isopropyl, isobutyl , sec-butyl,
tert-butyl, isopentyl, neopentyl, and isohexyl. Examples of
C.sub.1-C.sub.8 alkyl groups include, but are not limited to,
methyl, propyl, pentyl, hexyl, heptyl, 3-methylhex-1-yl,
2,3-dimethylpent-2-yl, 3-ethylpent-1-yl, octyl, 2-methylhept-2-yl,
2,3-dimethylhex-1-yl, and 2,3,3-trimethylpent-1-yl. An alkyl group
can be unsubstituted or substituted with one or more of halogen,
NH.sub.2, (alkyl)NH, (alkyl)(alkyl)N--, --N(alkyl)C(O)(alkyl),
--NHC(O)(alkyl), --NHC(O)H, --(CO)NH.sub.2, --C(O)NH(alkyl),
--(CO)N(alkyl)(alkyl), CN, OH, alkoxy, alkyl, C(O)OH,
--(CO)O(alkyl), --C(O)(alkyl), aryl, heteroaryl, heterocyclyl,
cycloalkyl, haloalkyl, aminoalkyl, --OC(O)(alkyl),
carboxyamidoalkyl-, NO.sub.2, and alkyl-CN.
[0064] "Alkoxy" refers to the group R--O-- where R is an alkyl
group, as defined above. Exemplary C.sub.1-C.sub.6 alkoxy groups
include but are not limited to methoxy, ethoxy, n-propoxy,
1-propoxy, n-butoxy and t-butoxy. An alkoxy group can be
unsubstituted or substituted with one or more of halogen, OH,
alkoxy, NH.sub.2, (alkyl)amino-, di(alkyl)amino-,
(alkyl)C(O)N(C.sub.1-C.sub.3 alkyl)-, (alkyl)carboxyamIDO1-,
HC(O)NH--, H.sub.2NC(O)--, (alkyl)NHC(O)--, di(alkyl)NC(O)--, CN,
C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)--, aryl, heteroaryl,
cycloalkyl, haloalkyl, amino(C.sub.1-C.sub.6 alkyl)-,
(alkyl)carboxyl-, or carboxyamidoalkyl-.
[0065] Aryl refers to an aromatic 6 to 14 membered hydrocarbon
group. Examples of a C.sub.6-C.sub.14 aryl group include, but are
not limited to, phenyl, .alpha.-naphthyl, .beta.-naphthyl,
biphenyl, anthryl, tetrahydronaphthyl, fluorenyl, indanyl,
biphenylenyl, and acenanaphthyl. Examples of a C.sub.6-C.sub.10
aryl group include, but are not limited to, phenyl,
.alpha.-naphthyl, .beta.-naphthyl, biphenyl, and
tetrahydronaphthyl. An aryl group can be unsubstituted or
substituted with one or more of alkyl, halogen, haloalkyl, alkoxy,
haloalkoxy, OH, hydroxyalkyl, O(hydroxyalkyl), --O(alkyl)C(O)OH,
-(alkyl)(alkoxy)halogen, NH.sub.2, aminoalkyl-, dialkylamino-,
C(O)OH, --(CO)O(alkyl), --OC(O)(alkyl), --O(alkyl)N(alkyl)(alkyl),
N-alkylamido-, --(CO)NH.sub.2, (alkyl)amido-, NO.sub.2,
(aryl)alkyl, alkoxy, aryloxy, heteroaryloxy, (aryl)amino,
(alkoxy)carbonyl-, (alkyl)amido-, (alkyl)amino, aminoalkyl-,
alkylcarboxyl-, (alkyl)carboxyamido-, (aryl)alkyl-, (aryl)amino-,
cycloalkenyl, di(alkyl)amino-, heteroaryl, (heteroaryl)alkyl-,
heterocyclyl, --O(heterocyclyl), heterocyclyl(alkyl)-,
(hydroxyalkyl)NH--, (hydroxyalkyl).sub.2N, --SO.sub.2(alkyl),
--NHC(O)(aryl), --C(O)NH(aryl), --NHC(O)(heteroaryl),
--(CO)NH(heteroaryl) or a spiro substituent.
[0066] The term "bicycle" or "bicyclic" as used herein refers to a
molecule that features two fused rings, which rings are a
cycloalkyl, heterocyclyl, or heteroaryl. In one embodiment, the
rings are fused across a bond between two atoms. The bicyclic
moiety formed therefrom shares a bond between the rings. In another
embodiment, the bicyclic moiety is formed by the fusion of two
rings across a sequence of atoms of the rings to form a bridgehead.
Similarly, a "bridge" is an unbranched chain of one or more atoms
connecting two bridgeheads in a polycyclic compound. In another
embodiment, the bicyclic molecule is a "Spiro" or "spirocyclic"
moiety. The spirocyclic group is a carbocyclic or heterocyclic ring
which bound through a single carbon atom of the spirocyclic moiety
to a single carbon atom of a carbocyclic or heterocyclic moiety. In
one embodiment, the spirocyclic group is a cycloalkyl and is bound
to another cycloalkyl. In another embodiment, the spirocyclic group
is a cycloalkyl and is bound to a heterocyclyl. In a further
embodiment, the spirocyclic group is a heterocyclyl and is bound to
another heterocyclyl. In still another embodiment, the spirocyclic
group is a heterocyclyl and is bound to a cycloalkyl.
[0067] "(Aryl)alkyl" refers to an alkyl group, as defined above,
wherein one or more of the alkyl group's hydrogen atoms has been
replaced with an awl group as defined above. (C.sub.6-C.sub.14
aryl)alkyl-moieties include benzyl, benzhydryl, 1-phenylethyl,
2-phenylethyl, 3-phenylpropyl, 2-phenylpropyl, 1-naphthylmethyl,
2-naphthylmethyl and the like. An (aryl)alkyl group can be
unsubstituted or substituted with one or more of halogen, CN,
NH.sub.2, OH, (alkyl)amino-, di(alkyl)amino-, (alkyl)C(O)N(alkyl)-,
(alkyl)carboxyamido-, HC(O)NH--, H.sub.2NC(O)--, (alkyl)NHC(O)--,
di(alkyl)NC(O)--, CN, OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-,
(alkyl)C(O)--, aryl, heteroaryl, cycloalkyl, haloalkyl,
amino(alkyl)-, (alkyl)carboxyl-, carboxyamidoalkyl-, or
NO.sub.2.
[0068] "(Alkoxy)carbonyl-" refers to the group alkyl-O--C(O)--.
Exemplary (C.sub.1-C.sub.6 alkoxy)carbonyl- groups include but are
not limited to methoxy, ethoxy, n-propoxy, 1-propoxy, n-butoxy and
t-butoxy. An (alkoxy)carbonyl group can be unsubstituted or
substituted with one or more of halogen, OH, NH.sub.2,
(alkyl)amino-, di(alkyl)amino-, (alkyl)C(O)N(alkyl)-,
(alkyl)carboxyamido-, HC(O)NH--, H.sub.2NC(O)--, (alkyl)NHC(O)--,
di(alkyl)NC(O)--, CN, alkoxy, C(O)OH, (alkoxy)carbonyl-,
(alkyl)C(O)--, aryl, heteroaryl, cycloalkyl, haloalkyl,
amino(alkyl)-, (alkyl)carboxyl-, carboxyamidoalkyl-, or
NO.sub.2.
[0069] "(Alkyl)amido-" refers to a --C(O)NH-- group in which the
nitrogen atom of said group is attached to a C.sub.1-C.sub.6 alkyl
group, as defined above. Representative examples of a
(C.sub.1-C.sub.6 alkyl)amido- group include, but are not limited
to, --C(O)NHCH.sub.3, --C(O)NHCH.sub.2CH.sub.3,
--C(O)NHCH.sub.2CH.sub.2CH.sub.3,
--C(O)NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3, --C(O)NH
CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--C(O)NHCH(CH.sub.3).sub.2, --C(O)NHCH.sub.2CH(CH.sub.3).sub.2,
--C(O)NHCH(CH.sub.3)CH.sub.2CH.sub.3, --C(O)NH--C(CH.sub.3).sub.3
and --C(O)NHCH.sub.2C(CH.sub.3).sub.3.
[0070] "(Alkyl)amino-" refers to an --NH group, the nitrogen atom
of said group being attached to a alkyl group, as defined above.
Representative examples of an (C.sub.1-C.sub.6 alkyl)amino-group
include, but are not limited to CH.sub.3NH--, CH.sub.3CH.sub.2NH--,
CH.sub.3CH.sub.2CH.sub.2NH--, CH.sub.3CH.sub.2CH.sub.2CH.sub.2NH--,
(CH.sub.3).sub.2CHNH--, (CH.sub.3).sub.2CHCH.sub.2NH--,
CH.sub.3CH.sub.2CH(CH.sub.3)NH-- and (CH.sub.3).sub.3CNH--. An
(alkyl)amino group can be unsubstituted or substituted on the alkyl
moiety with one or more of halogen, NH.sub.2, (alkyl)amino-,
di(alkyl)amino-, (alkyl)C(O)N(alkyl)-, (alkyl)carboxyamido-,
HC(O)NH--, H.sub.2NC(O)--, (alkyl)NHC(O)--, di(alkyl)NC(O)--, CN,
OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)--, aryl,
heteroaryl, cycloalkyl, haloalkyl, amino(alkyl)-, (alkyl)carboxyl-,
carboxyamidoalkyl-, or NO.sub.2.
[0071] "Aminoalkyl-" refers to an alkyl group, as defined above,
wherein one or more of the alkyl group's hydrogen atoms has been
replaced with --NH.sub.2; one or both H of the NH.sub.2 may be
replaced by a substituent.
[0072] "Alkylcarboxyl-" refers to an alkyl group, defined above
that is attached to the parent structure through the oxygen atom of
a carboxyl (C(O)--O--) functionality. Examples of (C.sub.1-C.sub.6
alkyl)carboxyl- include acetoxy, propionoxy, propylcarboxyl, and
isopentylcarboxyl.
[0073] "(Alkyl)carboxyamido-" refers to a --NHC(O)-- group in which
the carbonyl carbon atom of said group is attached to a
C.sub.1-C.sub.6 alkyl group, as defined above. Representative
examples of a (C.sub.1-C.sub.6 alkyl)carboxyamido-group include,
but arc not limited to, --NHC(O)CH.sub.3, --NHC(O)CH.sub.2CH.sub.3,
--NHC(O)CH.sub.2CH.sub.2CH.sub.3,
--NHC(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--NHC(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--NHC(O)CH(CH.sub.3).sub.2, NHC(O)CH.sub.2CH(CH.sub.3).sub.2,
--NHC(O)CH(CH.sub.3)CH.sub.2CH.sub.3, --NHC(O)--C(CH.sub.3).sub.3
and --NHC(O)CH.sub.2C(CH.sub.3).sub.3.
[0074] "(Aryl)amino" refers to a radical of formula (aryl)-NH--,
wherein aryl is as defined above. "(Aryl)oxy" refers to the group
Ar--O-- where Ar is an aryl group, as defined above.
[0075] "Cycloalkyl" refers to a non-aromatic, saturated, partially
saturated, monocyclic, bicyclic or polycyclic hydrocarbon 3 to 12
membered ring system. Representative examples of a C.sub.3-C.sub.12
cycloalkyl include, but are not limited to, cyclopropyl,
cyclopentyl, cycloheptyl, cyclooctyl, decahydronaphthalen-1-yl,
octahydro-1H-inden-2-yl, decahydro-1H-benzo[7]annulen-2-yl, and
dodecahydros-indacen-4-yl. Representative examples of a
C.sub.3-C.sub.10 cycloalkyl include, but are not limited to,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl, decahydronaphthalen-1-yl, and octahydro-1H-inden-2-yl.
Representative examples of a C.sub.3-C.sub.8 cycloalkyl include,
but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, and octahydropentalen-2-yl. A
cycloalkyl can be unsubstituted or substituted with one or more of
halogen, NH.sub.2, (alkyl)NH, (alkyl)(alkyl)N--,
--N(alkyl)C(O)(alkyl), --NHC(O)(alkyl), --NHC(O)H, --C(O)NH.sub.2,
--C(O)NH(alkyl), --(CO)N(alkyl)(alkyl), CN, OH, alkoxy, alkyl,
C(O)OH, --(CO)O(alkyl), --(CO) alkyl), aryl, heteroaryl,
cycloalkyl, haloalkyl, aminoalkyl-, --OC(O)(alkyl),
carboxyamidoalkyl-, and NO.sub.2. Additionally, each of any two
hydrogen atoms on the same carbon atom of the carbocyclic ring can
be replaced by an oxygen atom to form an oxo (.dbd.O)
substituent.
[0076] "Halo" or "halogen" refers to --F, --Cl, --Br and --I.
[0077] "C.sub.1-C.sub.6 haloalkyl" refers to a C.sub.1-C.sub.6
alkyl group, as defined above, wherein one or more of the
C.sub.1-C.sub.6 alkyl group's hydrogen atoms has been replaced with
F, Cl, Br, or I. Each substitution can be independently selected
from F, Cl, Br, or I. Representative examples of an C.sub.1-C.sub.6
haloalkyl-group include, but are not limited to, --CH.sub.2F,
--CCl.sub.3, --CF.sub.3, CH.sub.2CF.sub.3, --CH.sub.2C.sub.1,
--CH.sub.2CH.sub.2Br, --CH.sub.2CH.sub.2I,
--CH.sub.2CH.sub.2CH.sub.2F, --CH.sub.2CH.sub.2CH.sub.2C.sub.1,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2Br,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2I,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2Br,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2I,
--CH.sub.2CH(Br)CH.sub.3, --CH.sub.2CH(Cl)CH.sub.2CH.sub.3,
--CH(F)CH.sub.2CH.sub.3 and --C(CH.sub.3).sub.2(CH.sub.2Cl).
[0078] "Heteroaryl" refers to a monocyclic, bicyclic, or polycyclic
aromatic ring system containing at least one ring atom selected
from the heteroatoms oxygen, sulfur and nitrogen. Examples of
C.sub.1-C.sub.9 heteroaryl groups include furan, thiophene, indole,
azaindole, oxazole, thiazole, isoxazole, isothiazole, imidazole,
N-methylimidazole, pyridine, pyrimidine, pyrazine, pyrrole,
N-methylpyrrole, pyrazole, N-methylpyrazole, 1,3,4-oxadiazole,
1,2,4-triazole, 1-methyl-1,2,4-triazole, 1H-tetrazole,
1-methyltetrazole, benzoxazole, benzothiazole, benzofuran,
benzisoxazole, benzimidazole, N-methylbenzimidazole,
azabenzimidazole, indazole, quinazoline, quinoline, and
isoquinoline. Bicyclic C.sub.1-C.sub.9 heteroaryl groups include
those where a phenyl, pyridine, pyrimidine or pyridazine ring is
fused to a 5 or 6-membered monocyclic heteroaryl ring having one or
two nitrogen atoms in the ring, one nitrogen atom together with
either one oxygen or one sulfur atom in the ring, or one O or S
ring atom. Examples of monocyclic C.sub.1-C.sub.4 heteroaryl groups
include 2H-tetrazole, 3H-1,2,4-triazole, furan, thiophene, oxazole,
thiazole, isoxazole, isothiazole, imidazole, and pyrrole. A
heteroaryl group can be unsubstituted or substituted with one or
more of C.sub.1-C.sub.6 alkyl, halogen, haloalkyl, OH, CN,
hydroxyalkyl, NH.sub.2, aminoalkyl-, dialkylamino-, C(O)OH,
--(CO)O-(alkyl), --OC(O)(alkyl), N-alkylamido-, --(CO)NH.sub.2,
(alkyl)amido-, --NO.sub.2, (aryl)alkyl, alkoxy, aryloxy,
heteroaryloxy, (aryl)amino, (alkoxy)carbonyl-, (alkyl)amido-,
(alkyl)amino, aminoalkyl-, alkylcarboxyl-, (alkyl)carboxyamido-,
(aryl)alkyl-, (aryl)amino-, cycloalkenyl, di(alkyl)amino-,
heteroaryl, (heteroaryl)alkyl-, heterocyclyl, heterocyclyl(alkyl)-,
(hydroxyalkyl)NH--, (hydroxyalkyl).sub.2N, --NHC(O)aryl,
--(CO)NHaryl, --NHC(O)heteroaryl, --(CO)NH(heteroaryl), or a Spiro
substituent.
[0079] "Heterocycle" or "heterocyclyl" refers to monocyclic,
bicyclic, polycyclic, or bridged head molecules in which at least
one ring atom is a heteroatom. A heterocycle may be saturated or
partially saturated. Exemplary C.sub.1-C.sub.9 heterocyclyl groups
include but are not limited to aziridine, oxirane, oxirene,
thiirane, pyrroline, pyrrolidine, dihydrofuran, tetrahydrofuran,
dihydrothiophene, tetrahydrothiophene, dithiolane, piperidine,
1,2,3,6-tetrahydropyridine-1-yl, tetrahydropyran, pyran, thiane,
thiine, piperazine, azepane, diazepane, oxazine,
5,6-dihydro-4H-1,3-oxazin-2-yl, 2,5-diazabicyclo[2.2.1]heptane,
2,5-diazabicyclo[2.2.2]octane, 3,6-diazabicyclo[3.1.1]heptane,
3,8-diazabicyclo[3.2.1]octane, 6-oxa-3,8-diazabicyclo[3.2.1]octane,
7-oxa-2,5-diazabicyclo[2.2.2 ]octane,
2,7-dioxa-5-azabicyclo[2.2.2]octane,
2-oxa-5-azabicyclo[2.2.1]heptane-5-yl,
2-oxa-5-azabicyclo[2.2.2]octane,
3,6-dioxa-8-azabicyclo[3.2.1]octane,
3-oxa-6-azabicyclo[3.1.1]heptane,
3-oxa-8-azabicyclo[3.2.1]octan-8-yl,
5,7-dioxa-2-azabicyclo[2.2.2]octane,
6,8-dioxa-3-azabicyclo[3.2.1]octane,
6-oxa-3-azabicyclo[3.1.1]heptane,
8-oxa-3-azabicyclo[3.2.1]octan-3-yl,
2-methyl-2,5-diazabicyclo[2.2.1]heptane-5-yl,
1,3,3-trimethyl-6-azabicyclo[3.2.1]oct-6-yl,
3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl-,
7-methyl-3-oxa-7,9-diazabicyclo[3.3.1]nonan-9-yl,
9-oxa-3-azabicyclo[3.3.1]nonan-3-yl,
3-oxa-9-azabicyclo[3.3.1]nonan-9-yl,
3,7-dioxa-9-azabicyclo[3.3.1]nonan-9-yl,
4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl, thiazine, dithiane,
and dioxane. The contemplated heterocycle rings or ring systems
have a minimum of 3 members. Therefore, for example, C.sub.1
heterocyclyl radicals would include but are not limited to
oxaziranyl, diaziridinyl, and diazirinyl, C.sub.2 heterocyclyl
radicals include but are not limited to aziridinyl, oxiranyl, and
diazetidinyl, C.sub.9 heterocyclyl radicals include but are not
limited to azecanyl, tetrahydroquinolinyl, and
perhydroisoquinolinyl. A heterocyclyl group can be unsubstituted or
substituted with one or more of alkyl, halogen, alkoxy, haloalkyl,
OH, hydroxyalkyl, --(CO)-(hydroxyalkyl), NH.sub.2, aminoalkyl-,
dialkylamino-, C(O)OH, --(CO)O-(alkyl), --OC(O)(alkyl),
N-alkylamido-, --(CO)NH.sub.2, (alkyl)amido-, --(CO)-(alkyl)-CN,
(alkyl)-CN, or NO.sub.2.
[0080] "Heterocyclyl(alkyl)-" refers to an alkyl group, as defined
above, wherein one or more of the alkyl group's hydrogen atoms has
been replaced with a heterocycle group as defined above.
Heterocyclyl(C.sub.1-C.sub.6 alkyl)-moieties include
1-piperazinylethyl, 4-morpholinylpropyl, 6-piperazinylhexyl, and
the like. A heterocyclyl(alkyl) group can be unsubstituted or
substituted with one or more of halogen, NH.sub.2, (alkyl)amino-,
di(alkyl)amino-, (alkyl)C(O)N(alkyl)-, (alkyl)carboxyamido-,
HC(O)NH--, H.sub.2NC(O)--, (alkyl)NHC(O)--, di(alkyl)NC(O)--, CN,
OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)--, 4- to
7-membered monocyclic heterocycle, aryl, heteroaryl, or
cycloalkyl.
[0081] "Heteroaryl(alkyl)" refers to a heteroaryl which is attached
to an alkyl group and the heteroaryl is defined above.
[0082] "Hydroxyalkyl" refers to a alkyl group, as defined above,
wherein one or more of the alkyl group's hydrogen atoms has been
replaced with OH groups. Examples of C.sub.1-C.sub.6 hydroxyalkyl
moieties include, for example, --CH.sub.2OH, --CH.sub.2CH.sub.2OH,
--CH.sub.2CH.sub.2CH.sub.2OH, --CH.sub.2CH(OH)CH.sub.2OH,
--CH.sub.2CH(OH)CH.sub.3, --CH(CH.sub.3)CH.sub.2OH and higher
homologs.
[0083] "Perfluoroalkyl-" refers to alkyl group, defined above,
having two or more fluorine atoms. Examples of a C.sub.1-C.sub.6
perfluoroalkyl- group include CF.sub.3, CH.sub.2CF.sub.3,
CF.sub.2CF.sub.3 and CH(CF.sub.3).sub.2. This may also be referred
to as mono or difluorine substituted alkyl group such as CHF.sub.2
or CH.sub.2F.
[0084] A "subject" is a mammal, including but not limited to a
human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or
non-human primate, such as a monkey, chimpanzee, baboon or gorilla.
In certain embodiments, the subject is a human. In certain
embodiments, the subject is a non-human animal. The terms
"individual," "patient," and "subject" are used interchangeably
herein.
[0085] "Effective amount" means the amount of a compound that when
administered to a subject, tissue, cell, living organism, is
sufficient to inhibit the kynurenine pathway or activity of IDO1
and/or IDO2 and/or TDO.
[0086] "Therapeutically effective amount" means the amount of a
compound that, when administered to a subject for treating a
disease, is sufficient to effect such treatment for the disease.
The "therapeutically effective amount" can vary depending on the
compound, the disease and its severity, and the age, weight, etc.,
of the subject to be treated.
[0087] As used herein, the terms "therapeutic," "therapeutic
agent," "medication" and "medicament" may be used interchangeably
throughout the specification.
[0088] A "chemotherapeutic agent" is a biological (large molecule)
or chemical (small molecule) compound useful in the treatment of
cancer, regardless of mechanism of action. Classes of
chemotherapeutic agents include, but are not limited to: alkylating
agents, antimetabolites, spindle poison plant alkaloids,
cytotoxic/antitumor antibiotics, topoisomerase inhibitors,
proteins, antibodies, photosensitizers, and kinase inhibitors.
Chemotherapeutic agents include compounds used in "targeted
therapy" and non-targeted conventional chemotherapy.
[0089] As used herein, the term "isotopic variant" or
"isotopically" or "radio-labeled" refers to a compound of the
invention where one or more atoms are replaced or substitutd by an
atom having an atomic mass or mass number different from the atomic
mass or mass number typically found in nature (i.e., naturally
occurring). For example, an "isotopic variant" of a compound or a
"radio-labeled" compound can contain one or more non-radioactive
isotopes, such as for example, Deuterium (.sup.2H or D), Tritium
(.sup.3H), Carbon 11 (.sup.11C) Carbon-13 (.sup.13C), Carbon-14
(.sup.14C), Nitrogen-15 (.sup.15N), Oxygen-15 (.sup.15O), Oxygen-17
(.sup.17O), Oxygen-18 (18O), Fluorine-18 (.sup.18F), Sulphur-35
(.sup.35S), Chlorine-36 (.sup.36Cl), Bromium-75 (.sup.75Br),
Bromium-76 (.sup.76Br), Bromium-77 (.sup.77Br), Bromium-82
(.sup.82Br), Iodine-123 (.sup.123I), Iodine-125 (.sup.125I),
Iodine-131 (.sup.131I), or the like. It will be understood that, in
a compound where such isotopic substitution is made, the following
atoms, where present, may vary, so that for example, any hydrogen
may be .sup.2H/D, any carbon may be .sup.13C, or any nitrogen may
be .sup.15N, and that the presence and placement of such atoms may
be determined by a person having the skill in the art. Likewise,
the invention may include the preparation of isotopic variants with
radioisotopes, in the instance for example, where the resulting
compounds may be used for drug and/or substrate tissue distribution
studies. The radioactive isotopes tritium, i.e., .sup.3H, and
carbon-14, i.e., .sup.14C, are particularly useful for this purpose
in view of their ease of incorporation and ready means of
detection. Further, compounds may be prepared that are substituted
with positron emitting isotopes, such as .sup.11C, .sup.18F,
.sup.15O and .sup.13N, and would be useful in Positron Emission
Topography (PET) studies for examining substrate receptor
occupancy. All isotopic variants of the compounds provided herein,
radioactive or not, arc intended to be encompassed within the scope
of the invention.
[0090] The term "prodrug" means compounds that are transformed in
vivo to yield a compound of the present invention. The
transformation can occur by various mechanisms, such as through
hydrolysis, oxidation, or reduction in blood. A good discussion of
the use of prodrugs is provided by T. Higuchi and W. Stella,
"Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S.
Symposium Series, and in Bioreversible Carriers in Drug Design, ed.
Edward B. Roche, American Pharmaceutical Association an Pergamon
Press, 1987.
[0091] A "metabolite" is a product produced through metabolism in
the body of a specified compound or salt thereof. Metabolites of a
compound may be identified using routine techniques known in the
art and their activities determined using tests such as those
described herein. Such products may result for example from
oxidation, reduction, hydrolysis, amidation, deamidation,
esterification deesterification, enzymatic cleavage, and the like,
of the administered compound.
[0092] "Salts" refers to derivatives of the disclosed compounds
wherein the parent compound is modified by converting an existing
acid or base moiety to its salt form. Examples of salts include,
but are not limited to mineral acid (such as HCl, HBr,
H.sub.2SO.sub.4) or organic acid (such as acetic acid, benzoic
acid, trifluoroacetic acid), salts of basic residues such as amines
(primary, secondary, and tertiary amines, substituted amines
including naturally occurring substituted amines, cyclic amines
(such as caffeine, arginine, diethylamine, N-ethyl piperidine,
histidine, glucamine, isopropylamine, lysine, morpholine, N-ethyl
morpholine, piperazine, piperidine, triethylamine,
disopropylethylamine and trimethylamine), organic amines, for
example, ethylamine, ethanolamine, triethanolamine or amino acids);
alkali (such as Li, Na, K, Mg, Ca) or organic (such as
trialkyammonium) salts of acidic residues such as carboxylic acids;
alkyl or arylalkyl pyridinium salts; and the like. The salts of the
present invention can be synthesized from the parent compound which
contains a basic or acidic moiety by conventional chemical methods.
Generally, such salts can be prepared by reacting the free acid or
base forms of these compounds with a stoichiometric amounts of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two; generally, non-aqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are preferred.
[0093] The phrase "pharmaceutically acceptable" indicates that the
substance composition must be compatible chemically and/or
toxicologically, with other ingredients comprising a formulation,
and/or the mammal being treated therewith.
[0094] Representative "pharmaceutically acceptable salts" as used
herein refers to a pharmaceutically acceptable organic or inorganic
salts of a compound of the invention and include but are not
limited to those of an acid or base. Lists of suitable salts are
found in Remington's Pharmaceutical Sciences, 17.sup.th ed., Mack
Publishing Company, Easton, Pa., 1985, p 1418 and Journal of
Pharmaceutical Science, 66, 2 (1977), each of which is incorporated
in its entirety. In one embodiment, the pharmaceutical salt is
selected from amount water-soluble and water-insoluble salts, such
as the acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate,
bitartrate, bromide, butyrate, calcium, chloride, choline, citrate,
edisylate (camphorsulfonate), formate, fumarate, gluconate,
glucuronate, glutamate, hydrobromide, hydrochloride, iodide,
isonicotinate, lactate, lauryl sulfate, malate, maleate, mandelate,
methylsufonate, mesylate, nitrate, oleate, oxalate, palmitatc,
pantothcnatc, phosphate, acid phosphate, potassium, propionate,
p-toluenesulfonate, salicylate, sodium, stearate, succinate,
sulfate and tannate salts. A pharmaceutically acceptable salt may
involve the inclusion of another molecule such as an acetate ion, a
succinate in or other counter ion. The counter ion may be any
organic or inorganic moiety that stabilizes the charges on the
parent compound. Furthermore, a pharmaceutically acceptable salt
can have multiple counter ions. Hence, a pharmaceutically
acceptable salt can have one or more charged atoms and/or one or
more counter ion.
[0095] A "solvate" refers to a physical association or complex of
one or more solvent molecules and a compound of the invention. The
compounds of the invention may exist in unsolvate as well as
solvated forms. Examples of solvents that form solvates include but
are not limited to water, isopropanol, ethanol, methanol, DMSO,
ethyl acetate, acetic acid, and ethanolamine. The term "hydrate"
refers to the complex where the solvent molecule is water. This
physical association involves varying degrees of ionic and covalent
bonding, including hydrogen bonding. In certain instances, the
solvate will be capable of isolation, for example when one or more
solvent molecules are incorporated in the crystal lattice of the
crystalline solid. Preparation of solvates is generally known for
example, M. Caira et al. J. Pharmaceutical Sci. 93(3), 601-611
(2004). Similar preparations of solvates, hemisolvates, hydrates,
and the like are described by E. C. van Tonder et al. AAPS
PharmSciTech, 5(1), article 12 (2004); and A. L. Bingham et al.
Chem. Commun., 603-604 (2001). A typical, non-limiting, process
involves dissolving a compound of the invention in desired amount
of the desired solvent (organic or water or mixtures thereof) at a
higher than ambient temperature, and cooling the solution at a rate
sufficient to form crystals which are then isolated by standard
methods. Analytical techniques such as, for example, I.R.
spectroscopy, show the present of the solvent (or water in the
crystal as solvate (or hydrate)).
[0096] The term "synergistic" as used herein refers to a
therapeutic combination which is more effective than the additive
effects of the two or more single agents. A determination of a
synergistic interaction between one or more compounds of the
invention, or a compound of the invention and one or more
therapeutic agent may be based on the results obtained from the
assays described herein. The combination therapy may provide
"synergy" and prove "synergistic" i.e., the effect achieved when
the active ingredients used together is greater than the sum of the
effects that results from using the compounds separately. A
synergistic effect may be attained wen the active ingredients are:
(1) co-formulated and administered or delivered simultaneously in a
combined, unit dosage formulation; (2) delivered by alternation or
in parallel as separate formulations; or (3) by some other regimen.
When delivered in alternation therapy, a synergistic effect may be
attained when the compounds are administered or delivered
sequentially, e.g., by different injections in separate syringes.
In general, during alternation therapy, effective dosages of two or
more active ingredients are administered together.
[0097] The terms "enhance" or "enhancing," as used herein, means to
increase or prolong either in potency or duration a desired effect.
Thus, in regard to enhancing the effect of therapeutic agents, the
term "enhancing" refers to the ability to increase or prolong,
either in potency or duration, the effect of other therapeutic
agents on a system. An "enhancing-effective amount," as used
herein, refers to an amount adequate to enhance the effect of
another therapeutic agent in a desired system.
[0098] Some compounds within the present invention may possess one
or more chiral centers, and in some embodiments, each center exists
in the R or S configuration. The present invention includes each
separate enantiomer of such compounds as well as mixtures of the
enantiomers. Where multiple chiral centers exist in compounds of
the present invention, the invention includes each possible
combination of chiral centers within a compound, as well as all
possible enantiomeric and diastereomeric mixtures thereof. All
chiral, diastereomeric, and racemic forms of a structure are
intended, unless the specific stereochemistry or isomeric form is
specifically indicated. It is well known in the art how to prepare
optically active forms, such as by resolution of racemic forms or
by synthesis from optically active starting materials. Compounds of
the invention also include tautomeric forms. Tautomeric forms
result from the swapping of a single bond with an adjacent double
bond together the concomitant migration of a proton. For example,
enols and ketones are tautomers because they are rapidly
interconverted by treatment with either acid or base. Another
example of tautomerism is the aci- and nitro- forms of
phenylnitromethane, which are likewise formed by treatment with
acid or base. Tautomeric forms may be relevant to the attainment of
the optimal chemical reactivity and biological activity of a
compound of interest.
[0099] As used herein, the term "isotopic variant" refers to a
compound that contains unnatural proportions of isotopes at one or
more of the atoms that constitute such compound. For example, an
"isotopic variant" of a compound can contain one or more
non-radioactive isotopes, such as for example, deuterium (.sup.2H
or .sup.2D), carbon 13 (.sup.13C), nitrogen-15 (.sup.15N), or the
like. It will be understood that, in a compound where such isotopic
substitution is made, the following atoms, where present, may vary,
so that for example, any hydrogen may be .sup.2H/D, any carbon may
be .sup.13C, or any nitrogen may be .sup.15N, and that the presence
and placement of such atoms may be determined within the skill of
the art. Likewise, the invention may include the preparation of
isotopic variants with radioisotopes, in the instance for example,
where the resulting compounds may be used for drug and/or substrate
tissue distribution studies. The radioactive isotopes tritium,
i.e., .sup.3H, and carbon-14, i.e., .sup.14C, are particularly
useful for this purpose in view of their ease of incorporation and
ready means of detection. Further, compounds may be prepared that
are substituted with positron emitting isotopes, such as tic,
.sup.18F, .sup.15O and .sup.13N, and would be useful in Positron
Emission Topography (PET) studies for examining substrate receptor
occupancy. All isotopic variants of the compounds provided herein,
radioactive or not, are intended to be encompassed within the scope
of the invention.
[0100] For the purposes of the present disclosure, the terms
"compound," "compound of the invention," "test compounds," and
"composition of matter" stand equally well for inhibitors of
kynurenine pathway- and/or the IDO1- and/or IDO2- and/or TDO and
metabolites thereof described herein including all enantiomeric
forms, diastereomeric forms, racemic forms, racemic-diastereomeric
mixtures, optical isomers, tautomeric forms, salts, polymorphs, and
the like. The terms "compound," "compound of the invention," "test
compounds," and "composition of the matter" are used
interchangeably throughout the present specification.
[0101] For the purposes of the present disclosure, the terms
"disease," "condition," and "disorder" stand equally well for
conditions where a subject may benefit from regulation of
kynurenine pathway and/or IDO1, and/or IDO2, and/or TDO, and may be
used interchangeably throughout the present specification.
[0102] The following abbreviations are used and have the indicated
definitions:
TABLE-US-00001 Abbreviation Definition TMSCN trimethylsilyl nitrile
TFE trifluoro ethanol n-BuLi n-butyl lithium TMEDA tetramethylene
diamine DMF dimethylformamide DCM dichloromethane LDA lithium
diisopropylamide HCl hydrochloric acid NBS N-bromosuccinimide
Bis-Pin Bis(pinacolato) ACN or MeCN acetonitrile THF
tetrahydrofuran RT or rt room temperature DIPEA
N,N-diisopropylethylamine IPA isopropylamine BINAP
2,2'-bis(diphenylphosphino)-1,1'-binaphthyl DMAP 4-dimethylamino
pyridine DPPA diphenylphosphoryl azide PPA phenylpropanol amine TEA
triethylamine SEM trimethylsilylethoxy methyl Pd(OAc).sub.2
palladium acetate DACM Dimethyl acetamide PG Polypropyl glycol DOXO
Doxorubicin mpk Mg/Kg TDO tryptophan 2,3-dioxygenase
Pd(dppf)Cl.sub.2.cndot.DCM [1,1'Bis(diphenylphosphino)ferrocene]-
dichloropalladium(II), complex with dichloromethane SEMCl
2-(Trimethylsilyl)ethoxymethyl chloride MOMCl Methoxymethyl
chloride Dppf bis(diphenylphosphino)ferrocene MTBE methyl tertiary
butyl ether HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid ATCC American Type Culture Collection DMEM Dulbecco's Modified
Eagle Medium TCA trichloro acetic acid hIDO human indoleamine
2,3-dioxygenase CO.sub.2 carbon dioxide IFN.gamma. gamma-interferon
DMSO dimethylsulfoxide LTMS lithium trimethylsilyl
Pd(PPh.sub.3).sub.4 tetrakis(triphenylphosphine) palladium (0) TMS
trimethylsilane Pd.sub.2(dba).sub.3 tris(dibenzylideneacetone)
palladium (0) Boc butoxycarbonyl TFA trifluoroacetic acid HMPA
hexamethylphosphoramide MOM methoxymethyl ether LTMP lithium
2,2,6,6-tetramethylpiperidide TMP 2,2,6,6,-tetramethylpiperidine
XPhos 2-dicyclohexyl phosphino-2',4',6'-triisopropylbiphenyl
Ni(dppp)Cl.sub.2 dichloro(1,3-bis(diphenylphosphine) propane nickel
PEG Polyethylene Glycol PBS Phosphate buffered saline KYN
Kynurenine IDO indoleamine 2,3-dioxygenase TFA Trifluoroacetic acid
LHMDS Lithium Hexamethyldisilazide TMSOTf Trimethylsilyl trifluoro
methanesulfonate PPA Polyphosphoric acid EtOH Ethanol CrEL
Cremophore Ethanol
[0103] The invention provides compounds of formula (I) and
metabolites thereof, or pharmaceutically acceptable salts or
prodrugs thereof, and metabolites thereof, and pharmaceutical
composition thereof (collectively "compounds of the invention," or
"compounds," "test compounds," or "composition of matter"), which
are capable of reducing or eliminating immune-mediated disorders as
standalone therapy (monotherapy) or in combination with other
therapies, incuding without limitation, antiviral therapy,
anti-inflammation therapy, conventional chemotherapy, or in
combination with anti-cancer vaccines or in combination with
hormonal therapy to slow or prevent various conditions or diseases
including tumor growth. The invention further provides compounds
and compositions which function by decreasing levels of kynurenine
and/or altering the levels of tryptophan in plasma and/or tissues
through the inhibition of the enzymes indoleamine 2,3-dioxygenase-1
(IDO1) or indoleamine 2,3-dioxygenase-2 (IDO2) or tryptophan
2,3-dioxygenase (TDO) or any combination of the three enzymes.
[0104] In one embodiment, the compound is of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof.
##STR00006##
[0105] In this compound, X.sup.1 is CR.sup.1, N, or NO; X.sup.2 is
CR.sup.2, N, or NO; X.sup.3 is CR.sup.3, N, or NO; X.sup.4 is
CR.sup.4, N, or NO. In one embodiment, one or two of X.sup.1,
X.sup.2, X.sup.3 and X.sup.4 is N. In another embodiment, X.sup.1
is CR.sup.1, X.sup.2 is CR.sup.2, X.sup.3 is CR.sup.3, and X.sup.4
is CR.sup.4.
[0106] Y is O, S, or NR.sup.8 and Z is OR.sup.5, SR.sup.5 or
NR'R.sup.6.
[0107] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from the group consisting of H, optionally substituted
C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.2-C.sub.6
alkenyl, optionally substituted C.sub.2-C.sub.6 alkynyl, optionally
substituted C.sub.1-C.sub.6 alkoxy, mono or bicyclic optionally
substituted C.sub.6-C.sub.14 aryl, mono or bicyclic optionally
substituted heteroaryl, optionally substituted (aryl)alkyl,
(alkoxy)carbonyl, (alkyl)amido, (alkyl)amino, optionally
substituted mono or bicyclic cycloalkyl, optionally substituted
mono or bicyclic heterocyclyl, aminoalkyl, alkylcarboxyl,
(alkyl)carboxyamido, optionally substituted (aryl)amino, hydroxyl,
halogen, C.sub.1-C.sub.6 haloalkyl, optionally substituted
heterocyclyl(alkyl)-, optionally substituted heteroaryl(alkyl),
hydroxyalkyl, perfluoroalkyl, optionally substituted aryloxy,
optionally substituted heteroaryloxy, optionally substituted
C.sub.3-C.sub.8 cycloalkoxy, N(R.sup.7).sub.2, CN, NO.sub.2,
CO.sub.2H, CONR.sup.AR.sup.B, S(O).sub.nR.sup.7, and optionally
substituted heterocyclyloxy having 1 to 2 heteroatoms selected from
the group consisting of O, S(O).sub.n, and NR.sup.7.
[0108] In one embodiment, R.sup.1 is H, halogen, CN,
C.sub.1-C.sub.6 hydroxyalkyl, C.sub.1-C.sub.6 alkoxy, or
C.sub.1-C.sub.6 alkyl. In another embodiment, R.sup.1 is H. In yet
another embodiment, the R.sup.1 is a halogen. In still another
embodiment, R.sup.1 is a Cl. In yet another embodiment, R.sup.1 is
a methoxy or a methyl. In still another embodiment, R.sup.1 is
CN.
[0109] In a further embodiment, R.sup.2 is H, halogen, hydroxyl,
CN, N(R.sup.7).sub.2, mono or bicyclic optionally substituted
C.sub.6-C.sub.14 aryl, optionally substituted C.sub.1-C.sub.6
alkoxy, optionally substitutd C.sub.1-C.sub.6 alkyl, or optionally
substituted aryloxy. In a still further embodiment, R.sup.2 is F,
Cl, Br, or I. In yet another embodiment, R.sup.2 is H or optionally
substituted C.sub.1-C.sub.6 alkyl. In yet another embodiment,
R.sup.2 is optionally substituted C alkoxy or optionally
substituted aryloxy. In still another embodiment, R.sup.2 is
N(R.sup.7).sub.2 or mono or bicyclic optionally substituted
C.sub.6-C.sub.14 aryl.
[0110] In another embodiment, R.sup.3 is H, halogen, NO.sub.2 or
CN. In still a further embodiment, R.sup.3 is H. In yet another
embodiment, R.sup.3 is NO.sub.2 or CN.
[0111] H, optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted C.sub.2-C.sub.6 alkenyl, optionally substituted
C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.1-C.sub.6
alkoxy, mono or bicyclic optionally substituted C.sub.6-C.sub.14
aryl, CH.sub.2-aryl, mono or bicyclic optionally substituted
heteroaryl, optionally substituted (aryl)alkyl, (alkoxy)carbonyl,
(alkyl)amido, (alkyl)amino, optionally substituted mono or bicyclic
cycloalkyl, optionally substituted mono or bicyclic heterocyclyl,
aminoalkyl, alkylcarboxyl, (alkyl)carboxyamido, optionally
substituted (aryl)amino, hydroxyl, halogen, C.sub.1-C.sub.6
haloalkyl, optionally substituted heterocyclyl(alkyl)-, optionally
substituted heteroaryl(alkyl), hydroxyalkyl, perfluoroalkyl,
optionally substituted aryloxy, optionally substituted
heteroaryloxy, optionally substituted C.sub.3-C.sub.8 cycloalkoxy,
N(R.sup.7).sub.2, CN, NO.sub.2, CO.sub.2H, CONRARB, S(O)nR.sup.7,
and optionally substituted heterocyclyloxy having 1 to 2
heteroatoms selected from the group consisting of O, S(O)n, and
NR.sup.7, and n is 0 to 2.
[0112] In yet a further embodiment, R.sup.4 is H, halogen or CN. In
still another embodiment, R.sup.4 is optionally substituted phenyl.
In a further embodiment, R.sup.4 is phenyl substituted with one or
more C.sub.1-C.sub.6 alkoxy or halogen. In a further embodiment,
R.sup.4 is phenyl substituted with F, Cl, Br or 1.
[0113] In another embodiment, R.sup.4 is optionally substituted
alkyl, optionally substituted cycloalkyl, or optionally substituted
arylalkyl. In still another embodiment, R.sup.4 is
N(R.sup.7).sub.2. In yet another embodiment, R.sup.4 is optionally
substituted arylalkenyl or optionally substituted arylalkynyl. In
still another embodiment, R.sup.4 is optionally substituted
diarylamine or optionally substituted diphenylamine. In a further
embodiment, R.sup.4 is optionally substituted aryl, optionally
substituted bicylic aryl, heteroaryl, optionally substituted
heteroaryl, or bicyclic heteroaryl. In a still further embodiment,
R.sup.4 is an optionally substituted heterocyclyl.
[0114] In another embodiment, R.sup.4 is optionally substituted
pyridine, optionally substituted picolyl, optionally substituted
picolinamide. In yet another embodiment, R.sup.4 is R.sup.4 is
optionally substituted (alkyl)carboxyamido, (aryl)carboxyamido,
(alkyl)amido, alkylcarboxyl, (alkoxy)carbonyl, COOH,
C.sub.1-C.sub.6 cyclyloxy, heterocyclyloxy, aryloxy, heteroaryloxy,
perfluoroalkyl, S(O).sub.nN(R.sup.7).sub.2, or pyrimidine.
[0115] R.sup.A and R.sup.B are independently selected from among H,
optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted mono or bicyclic C.sub.6-C.sub.14 aryl, optionally
substituted mono or bicyclic heteroaryl, optionally substituted
(aryl)alkyl, optionally substituted mono or bicyclic
C.sub.3-C.sub.8 cycloalkyl, optionally substituted mono or bicyclic
heterocyclyl, C.sub.1-C.sub.6 haloalkyl, optionally substituted
heterocyclyl(alkyl), optionally substituted heteroaryl(alkyl),
hydroxyalkyl, and perfluoroalkyl.
[0116] In this compound, n is 0 to 2. In one embodiment, n is 0. In
another embodiment, n is 1. In a further embodiment, n is 2.
[0117] R.sup.7 is H, C.sub.1-C.sub.6 alkyl, mono or bicyclic
C.sub.6-C.sub.14 aryl, mono or bicyclic heteroaryl, (aryl)alkyl,
(alkoxy)carbonyl, (alkyl)amido, (alkyl)amino, mono or bicyclic
cycloalkyl, mono or bicyclic heterocyclyl, alkylcarboxyl,
heterocyclyl(alkyl), heteroaryl(alkyl), hydroxyalkyl,
perfluoroalkyl, aryloxy, heteroaryloxy, C.sub.3-C.sub.6
cycloalkoxy, or heterocyclyloxy having 1 to 2 heteroatoms selected
from the group consisting of O, S(O).sub.n, and NR.sup.A. R.sup.A
is H, C.sub.1-C.sub.6 alkyl, mono or bicyclic C.sub.6-C.sub.14
aryl, mono or bicyclic heteroaryl, (aryl)alkyl, (alkoxy)carbonyl,
(alkyl)amido, (alkyl)amino, mono or bicyclic cycloalkyl, mono or
bicyclic heterocyclyl, alkylcarboxyl, heterocyclyl(alkyl),
heteroaryl(alkyl), hydroxyalkyl, perfluoroalkyl, aryloxy,
heteroaryloxy, C.sub.3-C.sub.6 cycloalkoxy, or optionally
substituted heterocyclyloxy.
[0118] R.sup.8 is H or optionally substituted C.sub.1-C.sub.6
alkyl.
[0119] R.sup.5 and R.sup.6 are independently selected from among H,
optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted mono or bicyclic C.sub.6-C.sub.14 aryl, optionally
substituted mono or bicyclic heteroaryl, optionally substituted
(aryl)alkyl, optionally substituted mono or bicyclic cycloalkyl,
optionally substituted mono or bicyclic heterocyclyl,
C.sub.1-C.sub.6 haloalkyl, optionally substituted
heterocyclyl(alkyl), optionally substituted heteroaryl(alkyl),
hydroxyalkyl, and perfluoroalkyl. In one embodiment, R.sup.5 or
R.sup.6 is optionally substituted phenyl. In another embodiment,
R.sup.5 or R.sup.6 is of the following structure, wherein, R.sup.C
to R.sup.G are independently selected from among H, halogen,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy, heterocycle,
optionally substituted C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, CN, --O(aryl), C.sub.2-C.sub.6 alkynyl,
C(O)C.sub.1-C.sub.6 alkyl, --O--C.sub.1-C.sub.6 haloalkyl, and
optionally substituted aryl.
##STR00007##
[0120] In a further embodiment, R.sup.5 or R.sup.6 is of the
following structure, wherein, R.sup.C to R.sup.G are independently
selected from among H, halogen, CHF.sub.2, C(CH.sub.3)F.sub.2,
OCF.sub.3, OCH.sub.3, OCH(CH.sub.3).sub.2, morpholine, piperidine,
CH.sub.3, C(CH.sub.3).sub.3, CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2,
cyclopropyl, cyclohexyl, CH.sub.2-cyclopropyl, CH.sub.2-cyclobutyl,
benzyl, CN, phenoxy, ethynyl, C(O)CH.sub.3, and phenyl.
##STR00008##
[0121] In yet another embodiment, R.sup.5 or R.sup.6 is of the
following structure, wherein, R.sup.C to R.sup.G are independently
selected from the group consisting of H and optionally substituted
aryl. In one embodiment, R.sup.C to R.sup.G are independently
selected from among H and aryl substituted with one or more
halogen. In yet another embodiment, each halogen is independently
selected from F, Cl, Br, or I. In another embodiment, R.sup.C to
R.sup.G arc independently selected from among H and aryl
substituted with one or more Cl or F.
##STR00009##
[0122] In still a further embodiment, R.sup.5 or R.sup.6 is phenyl,
2-Br-4-F-phenyl, 2,3,4-tri-Cl-phenyl, 2,3-di-Cl-4-F-phenyl,
2,4-di-Cl-phenyl, 2-Cl-4-F-phenyl, 2-Cl-phenyl, 2-Et-phenyl,
2,4-di-F-3-Cl-phenyl, 2-F-3-CN-phenyl, 2,4-di-F-phenyl, 2-tetralin,
2,3-di-Me-phenyl, 2-Me-4-Br-phenyl, 2,4-di-Me-phenyl,
2,4-di-OMe-phenyl, 2-piperidine-phenyl, 3-Br-4,5-di-F-phenyl,
3-Br-4-F-phenyl, 3-Br-4-Me-phenyl, 3-Br-phenyl,
3-acetylene-4-F-phenyl, 3-acetylene-phenyl,
3-CF.sub.2Me-4-F-phenyl, 3-CF.sub.3-4-Br-phenyl,
3-CF.sub.3-4-Cl-phenyl, 3-CF.sub.3-4-F-phenyl, 3-CF.sub.3-phenyl,
3-CH.sub.2-cyclobutyl-phenyl, 3-CH.sub.2-cyclopropyl-phenyl,
3-CH.sub.2Ph-phenyl, 3-CHF2-phenyl, 3,4-di-Cl-phenyl,
3,5-di-Cl-4-F-phenyl, 3-Cl-4,6-di-F-phenyl, 3-Cl-4-F-phenyl,
3-Cl-4-I-phenyl, 3-Cl-4-Me-phenyl, 3-Cl-5-Me-phenyl,
3,6-di-Cl-phenyl, 3-Cl-6-F-phenyl, 3-Cl-6-OMe-phenyl, 3-Cl-phenyl,
3-CN-phenyl, 3-cyclohexyl-phenyl, 3-cyclopropyl-phenyl,
3-Et-phenyl, 3,4,6-tri-F-phenyl, 3,4-di-F-phenyl, 3,5-di-F-phenyl,
3-F-phenyl, 3-tetralin, 3-I-phenyl, 3-iPr-phenyl, 3-Me-4-Cl-phenyl,
3-Me-4-F-phenyl, 3,4-di-Me-phenyl, 3,5-di-Me-phenyl, 3-Me-phenyl,
3-OCF.sub.3-4-F-phenyl, benzo[d]dioxolane, 3-OiPr-phenyl,
3-OMe-4-Cl-phenyl, 3,5-di-OMe-phenyl, 3-OMe-phenyl, 3-Ph-phenyl,
3-chloropyridyl, 3-pyridyl, 3,5-di-tBu-phenyl, 3-tBu-phenyl,
4-Br-phenyl, 4-acetylene-phenyl, 4-CF.sub.3-phenyl,
4-CH.sub.2Ph-phenyl, 4-Cl-phenyl, 4-CN-phenyl, 4-COMe-phenyl,
4-F-phenyl, 4-Me-phenyl, 4-morpholine-phenyl, 4-OCF.sub.3-phenyl,
4-OPh-phenyl, or 5-Cl-6-F-phenyl.
[0123] In yet another embodiment, R.sup.5 or R.sup.6 is optionally
substituted heteroaryl.
[0124] In a further embodiment, R.sup.5 or R.sup.6 is pyridine
optionally substituted with one or more halogen. In one embodiment,
each halogen is independently selected from a group of F, Cl, Br
and I.
[0125] In one embodiment, R.sup.5 or R.sup.6 is pyrrolyl, indolyl,
pyrimidinyl, alkyl, benzyl, cycloalkylyl, heterocycloalkylyl, or
heterocycloalkylylalkyl.
[0126] In another embodiment, R.sup.5 or R.sup.6 is
benzo[d]dioxolane.
[0127] In yet a further embodiment, R.sup.5 or R.sup.6 is
tetrahydronaphthalene.
[0128] In another embodiment, the compound is of formula (I-A),
wherein X.sup.1-X.sup.4, Y, R.sup.5 and R.sup.6 are defined
herein.
##STR00010##
[0129] In a further embodiment, the compound is of formula (I-AA),
wherein X.sup.1-X.sup.4, R.sup.5 and R.sup.6 are defined
herein.
##STR00011##
[0130] In yet another embodiment, the compound is of formula
(I-AAA), wherein X.sup.1-X.sup.4, R.sup.5 and R.sup.6 are defined
herein.
##STR00012##
[0131] In still a further embodiment, the compound is of formula
(I-B), wherein X.sup.1-X.sup.4, Y, and R.sup.5 are defined
herein.
##STR00013##
[0132] In another embodiment, the compound is of formula (I-BB),
wherein X.sup.1-X.sup.4 and R.sup.5 are defined herein.
##STR00014##
[0133] In yet a further embodiment, the compound is of formula
(I-BBB), wherein X.sup.1-X.sup.4 and R.sup.5 are defined
herein.
##STR00015##
[0134] In still a further embodiment, the compound is of formula
(I-BBBB), wherein X.sup.1-X.sup.4, Y, and R.sup.5 are defined
herein.
##STR00016##
[0135] In another embodiment, the compound is of formula (1-BBBBB),
wherein X.sup.1-X.sup.4 and R.sup.5 are defined herein.
##STR00017##
[0136] In yet a further embodiment, the compound is of formula
(I-BBBBBB), wherein X.sup.1-X.sup.4 and R.sup.5 arc defined
herein.
##STR00018##
[0137] In still another embodiment, the compound is of formula
(I-C), wherein X.sup.1-X.sup.4 and R.sup.5 are defined herein.
##STR00019##
[0138] In a further embodiment, the compound is of formula (I-CC),
wherein X.sup.1-X.sup.4 and R.sup.5 are defined herein.
##STR00020##
[0139] In yet another embodiment, the compound is of formula (I-D),
wherein R.sup.5 and R.sup.6 arc defined herein.
##STR00021##
[0140] In still a further embodiment, the compound is of formula
(I-DD), wherein R.sup.5 and R.sup.6 are defined herein.
##STR00022##
[0141] In another embodiment, the compound is of formula (I-E),
wherein R.sup.5 and R.sup.6 are defined herein.
##STR00023##
[0142] In a further embodiment, the compound is of formula (I-EE),
wherein R.sup.5 and R.sup.6 are defined herein.
##STR00024##
[0143] In still another embodiment, the compound is of formula
(I-F), wherein R.sup.1, R.sup.2, and R.sup.4-R.sup.6 arc defined
herein.
##STR00025##
[0144] In yet a further embodiment, the compound is of formula
(I-FF), wherein R.sup.1, R.sup.2, and R.sup.4-R.sup.6 are defined
herein.
##STR00026##
[0145] In another embodiment, the compound is of formula (I-G),
wherein R.sup.1-R.sup.5 are defined herein.
##STR00027##
[0146] In still a further aspect, the compound is of formula
(I-GG), wherein R.sup.1-R.sup.5 are defined herein.
##STR00028##
[0147] Also falling within the scope of this invention are in vivo
metabolic products of the compounds of formula (I) described herein
and prodrugs thereof Such metabolic products may result from the
oxidation, reduction, hydrolysis, amidation, deamidation,
esterification, deesterification, enzymatic cleavage, and the like,
of the administered compound. Accordingly, compounds of the
invention include without limitation metabolites of compounds of
formula (I) and prodrugs thereof. Further, the invention includes
metabolites of compounds of formula (I), including compounds
produced synthetically and/or by a process comprising contacting a
compound of this invention with a mammal or a cell, for example, a
mammalian cell (including without limitation, rat, mice, human,
ape, monkey, rabbit, guinea pig, hamster, pig, cow, goat, sheep,
cat, dog etc.) or a eukaryotic cell such as a yeast cell, for a
period of time sufficient to yield a metabolic product thereof, and
prodrug thereof.
[0148] Metabolic products typically are identified by preparing a
radiolabeled (e.g., .sup.14C or .sup.3H) isotope of a compound of
the invention, administering it parentally in a detectable dose
(e.g., greater than about 0.5 mg/kg) to an animal such as a rat,
mouse, guinea pig, monkey, rabbit, bovine such a cow, ape, goat,
cat, or to human allowing sufficient time for metabolism to occur
(typically about 30 seconds to 30 hours) and isolating its
conversion products from urine, blood, or other biological samples.
These products are easily isolated since they are labeled (others
are isolated by the use of antibodies capable of binding eptitopes
surviving in the metabolite). Initial identification and analysis
of the metabolites, however, may also be performed using
non-radiolabeled compounds. In general, analysis of metabolites is
done in the same way as conventional drug metabolisms studies well
known to those skilled in the art. Metabolite structures are
determined in conventional fashion, e.g., MS, LC/MS/MS or NMR
analysis.
[0149] In one aspect, the invention relates to a purified and
isolated metabolite of the compounds of formula (I) and prodrugs
thereof, and pharmaceutically acceptable salts of the metabolites
thereof In another aspect, the invention relates to
pharmaceutically acceptable metabolites of compounds of formula (I)
and prodrugs thereof. In one aspect of the invention, metabolites
of compounds of formula (I) have a molecular mass less than that of
a corresponding or parent compound of formula (I). In another
aspect of the invention, the molecular mass of metabolites is
greater than that of corresponding or parent compound of formula
(I). In one embodiment of the invention, the molecular mass of
metabolites of compounds of formula (I) is less by 1 or 2 units
than that of corresponding or parent compound of formula (I). In
another embodimentof the invention, the molecular mass of
metabolites of compounds of formula (I) is less by 2 units. In yet
another aspect of the invention, metabolites are capable of
converting back to a corresponding (parent) compound in vitro and
in vivo. The conversion may be complete or partial. Accordingly,
metabolites of compounds of formula (I) are useful as prodrugs.
[0150] Compounds of formula (I) or metabolites thereof that are
basic in nature are capable of forming a wide variety of different
salts with various inorganic and organic acids. Although such salts
must be pharmaceutically acceptable for administration to animals
and mammals, it is often desirable in practice to initially isolate
a compounds or metabolite from a reaction mixture as a
pharmaceutically unacceptable salt and then simply convert the
latter back to a free base compound by treatment with an alkaline
reagent, and subsequently convert the free base to a
pharmaceutically acceptable acid addition salt. The acid addition
salts of the base compounds of this invention are readily prepared
by treating the base compound with a substantially equivalent
amount of a chosen mineral or organic acid in an aqueous solvent
medium or in a suitable organic solvent such as, but not limited
to, methanol or ethanol. Upon careful evaporation of the solvent,
the desired solid salt is obtained.
[0151] Acids which are used to prepare the pharmaceutically
acceptable acid addition salts of the base compounds of this
invention are those which form non-toxic acid addition salts, i.e.,
salts containing pharmacologically acceptable anions, such as
hydrochloride, hydrobromide, hydroiodide, nirate, sulfate or
bisulfate, phosphate or acid phosphate, acetate, lactate, citrate
or acid citrate, tartarate or bitartrate, succinate, maleate,
fumarate, gluconate, saccharate, benzoate, methanesulfonate and
palmoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphtoate)) salts
and the like.
[0152] Compounds of formula (I) or metabolites thereof which are
also acidic in nature, e.g., where R.sup.1-R.sup.6 includes a COOH
or tetrazole moiety, are capable of forming base salts with various
pharmacologically acceptable cations. Examples of such salts
include the alkali metal or alkaline-earth metal salts and
particularly, the sodium and potassium salts. These salts are
prepared by conventional techniques. The chemical bases which arc
used as reagents to prepare the pharmaceutically acceptable base
salts of this invention are those which form non-toxic base salts
with the herein described acidic metabolites of the compounds of
formula (I). These non-toxic base salts include those derived from
such pharmacologically acceptable cations as sodium, potassium,
calcium, and magnesium etc. These salts can easily be prepared by
treating the corresponding acidic compounds with an aqueous
solution containing the desired acceptable cations, and then
evaporating the resulting solution to dryness, preferably under
reduced pressure. Alternatively, they can also be prepared by
mixing lower alkanoic solutions of the acidic compounds and the
desired alkali metal alkoxide together, and then evaporating the
resulting solution to dryness in the same manner as before. In
either case, stoichiometric quantitities of reagents are preferably
employed in order to ensure completeness of reaction and maximum
product yields.
[0153] One aspect of the invention is directed to prodrugs of
compounds of the invention. In a further embodiment, a compound of
the invention may be a prodrug of a compound of formula (I).
Another aspect of the invention is directed to prodrugs of
metabolites of the compounds.
[0154] In a one embodiment, a compound of the invention may be a
prodrug of a compound of formula (I). Acetyl, amide, carbamate,
carbonate, ester or carbonate prodrugs of compounds of formula (I)
may be prepared using the methods described herein. In one
embodiment, a compound of formula (I) may be reacted with an acyl
chloride. In another embodiment, the acyl chloride may be
R.sup.ZC(O)Cl, where R.sup.Z is C.sub.1-C.sub.6 optionally
substituted alkyl, C.sub.6-C.sub.10 optionally substituted aryl, or
heteroaryl. In an embodiment, the prodrug is an acylated compound
of formula (I) which has the formula (II), wherein R.sup.9 is
optionally-substituted C.sub.1-C.sub.6 alkyl.
##STR00029##
[0155] In one embodiment, the prodrug is of formula (II) wherein
R.sup.9 is methyl. In a further embodiment, the reaction may be
performed in the presence of a base such as potassium
tert-butoxide, to provide a prodrug of compound of formula (I). In
a further embodiment, the reaction may be performed in the presence
of a base such as pyridine. As well, acetyl amide prodrugs of
compounds of formula (I) may be prepared by reaction of a compound
of formula (I) with MeCN. In one embodiment, the reaction is
performed under acidic conditions.
[0156] Thus, in one embodiment, the prodrug is an acetylated
compound of formula (I). In a further embodiment, the prodrug is a
phosphamide of the compound of formula (I). In still a further
embodiment, the prodrug is an acetate of the compound of formula
(I).
[0157] In another embodiment, the prodrug is a carbamate of the
compound of formula (I) which has the formula (III), wherein
R.sup.10 is optionally-substituted C.sub.1-C.sub.6 alkyl, aryl,
(aryl)alkyl, heteroaryl, or (heteroaryl)alkyl. In one embodiment,
the prodrug structure is formula (III) wherein R.sup.10 is selected
from methyl, ethyl, benzyl, (C.sub.1-C.sub.6 alkoxy)methyl, and
1-(C.sub.1-C.sub.6 alkoxy)ethyl.
##STR00030##
[0158] In yet another embodiment, the prodrug is a (bis)carbamate
of the compound of formula (I) which has the formula (IV), wherein
R.sup.11 is optionally-substituted C.sub.1-C.sub.6 alkyl, aryl,
(aryl)alkyl, heteroaryl, or (heteroaryl)alkyl. In one embodiment,
the prodrug structure is formula (IV) wherein R.sup.11 is selected
from methyl, ethyl, benzyl, (C.sub.1-C.sub.6 alkoxy)methyl, and
1-(C.sub.1-C.sub.6 alkoxy)ethyl.
##STR00031##
[0159] Thus, in still a further embodiment, the carbamate group is
selected from the following:
##STR00032##
[0160] Prodrugs of compounds of formula (I) may be prepared and
used as a means to modulate the pharmacokinetic properties, using
various methods known to those skilled in the art. See, e.g.,
Rautio, Nature Reviews Drug Discovery, 7:255-270 (2008) and
Ettmayer, J. Med. Chem., 47:2393-2404 (2004), which are hereby
incorporated by reference. In the case of drugs containing a
hydroxy moiety, acetyl and other ester analogs are contemplated for
use as prodrugs. See, e.g., Beaumont, Current Drug Metabolism,
4:461-485 (2003), which is hereby incorporated by reference. In the
case of drugs containing an amine moiety, prodrugs containing
amides and carbamates are contemplated. See, e.g., Simplicio,
Molecules, 13:519-547 (2008), which is hereby incorporated by
reference. As specific examples, (alkoxycarbonyloxy)alkyl
carbamates, (acyloxy)alkyl carbamates, and (oxodioxolenyl)alkyl
carbamates may be utilized as effective prodrug strategies for
amines. See, e.g., Li, Bioorg. Med. Chem. Lett., 7:2909-2912
(1997); Alexander, J. Med. Chem., 34:78-81 (1991); Alexander, J.
Med. Chem., 31:318-322 (1988); and Alexander, J. Med. Chem.,
39:480-486 (1996), all of which are incorporated by reference
herein.
[0161] Compounds of formula (I) and metabolites thereof, and
pharmaceutically acceptable salts and prodrugs thereof, as well as
metabolites of prodrugs of compounds of formula (I) (collectively
"compounds of the invention," "compounds" or "test compounds") are
well within the scope of this invention.
Schemes for Preparation
[0162] The compounds of the invention may be synthesized by
synthetic routes that include processes analogous to those
well-known in the chemical arts and those included in the present
application. Starting materials are generally available from
commercial sources such as Sigma Aldrich Chemicals (Milwakee, Wis.)
or are readily prepared using methods well known to those skilled
in the art (e.g., prepared by methods generally described in Louis
F. Fieser and Mary Fieser, Reagents for Organic Sythesis, v. 1-19,
Wiley, N.Y. (1967-1999 ed.), or Vogel's Textbook of Practical
Organic Chemistry (5th Edition) A. I. Vogel et al., or Beilsteins
Handbuch der organischen Chemi, 4, Aufl. Ed. Springer-Verlag,
Berlin, including supplements (also available via the Beilstein and
Reaxys online database).
[0163] Compounds of the invention may be converted into a
pharmaceutically acceptable salt, and a salt may be converted into
the free base compound, by conventional methods. Compounds of the
invention may be therapeutically effective as a free base or as a
pharmaceutically acceptable salt, depending on the desired
properties such as solubility, dissolution, hygroscopic nature, and
pharmacokinetics. Examples of pharmaceutically acceptable salts
include salts with inorganic acids, such as hydrochloric acids,
trifluoroacetic acid, propprionic acid, oxalic acid, malonic acid,
accinic acid, fumaric acid, maleic acid, lactic acid, malic acid,
tartaric acid, citric acid, ethanesulfonic acid, aspartic acid and
glutamic acid. The salt may be a mesylate, a hydrochloride, a
phosphate, a benzenesulfonate, or a sulfate. Salts may be
mono-salts or bis-salts. For example, the mesylate may be the
monomesylate or the bismesylate.
[0164] Compounds of the invention may also exist as hydrates or
solvates.
[0165] Protection of functional groups (e.g., primary or secondary
amines) of the intermediates may be necessary in preparing
compounds of formula (I)-(IV). The need for such protection will
vary depending on the nature of the remote functionality and the
conditions of the preparation methods. Suitable amino-protecting
groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (Boc),
benzyloxycarbonyl (CBz) and 9-fluorenylethyleneoxycarbonyl (Fmoc).
The need for such protection is readily determined by one skilled
in the art. For a general description of protection groups and
their use, see T.W. Greene, Protective groups in Organic Synthesis,
John Wiley & Sons, New York, 1991.
[0166] Methods useful for making the compounds of formula (I) are
set forth in the Examples below and generalized in Schemes 1-28.
One of skill in the art will recognize that Schemes 1-28 can be
adapted to produce other compounds of formula (I), prodrugs,
metabolites, and pharmaceutically acceptable salts of compounds of
formula (I) according to the present invention.
##STR00033##
[0167] Scheme 1 depicts a synthesis of fluoropyridin derivative
(I-C). Compound A1 was treated with an amine (R.sup.5--NH.sub.2).
In one embodiment, the amine (R.sup.5--NH.sub.2) was an optionally
substituted aniline. In another embodiment, the amine
(R.sup.5--NH.sub.2) was a heterocyclic amine. In yet another
embodiment, the amine (R.sup.5--NH.sub.2) was an alkylamine. After
the complete disappearance of starting materials, an alkylsilyl
cyanide was added to provide compound (I-C). In one embodiment, the
alkylsilyl cyanide was TMSCN.
##STR00034##
[0168] Scheme 1A depicts a synthesis of compound (I-H). 3-Hydroxy
pyridine-2-carboxaldehyde 1-1 was treated with an amine
(R.sup.5NH.sub.2). In one embodiment, the amine (R.sup.5--NH.sub.2)
is an optionally substituted aniline. After the complete
disappearance of starting materials, TMSCN was added to provide
compound (I-H).
##STR00035##
[0169] Scheme 1B depicts a synthesis of fluoropyridin derivative
1-2. 3-Hydroxy pyridine 2-carboxaldehyde 1-1 was treated with
3-chloro-4-fluoroaniline. After complete disappearance of starting
materials, TMSCN was added to provide compound 1-2.
##STR00036##
[0170] Scheme 2 provides the compounds of formula (I-C). A sodium
or potassium alkoxide or NaH was added to a solution of compound
B1. In one embodiment, the potassium alkoxide was potassium
tert-butoxide. Compound B1 was reacted with methoxymethyl chloride
or 2-(trimethylsilyl)ethoxymethyl chloride (SEMC1) to provide MOM
or SEM protected compound C1. TMEDA, HMPA, TEA, or DIPEA was then
added to a solution of compound C1, followed by addition of an
alkyllithium reagent and then DMF, N-formylpiperidine or
ethylformate to provide carbaldehyde D1. In one embodiment, the
alkyl-lithium reagent was n-BuLi. Deprotection of the MOM or SEM
group provided the 3-hydroxy carbaldehyde compound A1. Compound A1
was then treated with an amine (R.sup.5--NH.sub.2) in the presence
of an acid to provide imine E1. Imine E1 then underwent Strecker
reaction followed by intramolecular cyclization using TMSCN to form
fluoropyridin compound (I-C).
##STR00037##
[0171] Scheme 2A provides compounds of formula (I-I). Potassium
tert-butoxide was added to a solution of 3-hydroxypyridine 2-1.
Methoxymethyl chloride was then added to afford the desired MOM
protected compound 2-2. TMEDA was then added to compound 2-2.
n-BuLi was then added to this solution. DMF was then added to
provide the MOM protected carbaldehyde 2-3. Deprotection of the MOM
group provided compound 2-4. In one embodiment, the deprotection
was performed using 3N HCl/THF. Compound 2-4 was then treated with
an amine (R.sup.5--NH.sub.2) to provide imine E2 as the
intermediate. In one embodiment, the amine (R.sup.5--NH.sub.2) was
an aniline. Imine E2 was then reacted with TMSCN to form
fluoropyridin compound (I-I).
##STR00038##
[0172] Scheme 2B provides the formation of compound 2-6. Potassium
tert-butoxide was added to 3-hydroxypyridine 2-1. Methoxymethyl
chloride was then added to afford the desired MOM protected
compound 2-2. TMEDA was then added to compound 2-2 followed by
addition of n-BuLi. DMF was then added to give the MOM protected
carbaldehyde 2-3. Deprotection of the MOM group provided
3-hydroxypyridine-2-carbaldehyde 2-4. In one embodiment, the
deprotection was performed using 3N HCl. Compound 2-4 was treated
with 3-chloro-4-fluoroaniline to provide imine 2-5. The
corresponding imine underwent Strecker reaction followed by
intramolecular cyclization using TMSCN to form fluoropyridin
2-6.
##STR00039##
[0173] Scheme 3 shows the synthesis of compound (I-J). Compound F1
was coupled with an amine (R.sup.5--NH.sub.2). In one embodiment,
the amine (R.sup.5--NH.sub.2) was an optionally substituted aniline
to provide the corresponding imine compound G1. Compound G1 was
reacted with TMSCN to provide target compound (I-J).
##STR00040##
[0174] Scheme 3A shows the synthesis of compound (I-K). First,
compound F2 was coupled with an amine (R.sup.5--NH.sub.2) to
provide the corresponding G2 imine. Imine G2 was then reacted with
TMSCN to provide compound (I-K).
##STR00041##
[0175] Scheme 3B shows the synthesis of
[2-amino-3-(3-chloro-4-fluorophenyl)amino]-7-methyl
furo[2,3-c]pyridin-4-yl)methanol 3-3. Pyridoxal hydrochloride 3-1
was reacted with 4-fluoro-3-chloro aniline to provide corresponding
imine intermediate 3-2. Reaction with TMSCN provided compound
3-3.
##STR00042##
[0176] Scheme 4 depicts another synthesis of furopyridine
derivative (I-C). To a stirred solution of compound III was added a
potassium or sodium alkoxide to provide MOM protected compound C1.
In one embodiment, the potassium alkoxide was potassium
tert-butoxide. Methoxymethyl chloride was then added to form MOM
protected compound C1. LDA or LTMP was added to the solution of
compound C1, followed by N-formylpiperidine or DMF to provide
compound D1. Aldehyde D1 was deprotected via its reaction with an
acid. In one embodiment, the acid was HCl or TFA/DCM. Deprotected
aldehyde A1 was coupled with an amine (R.sup.5--NH.sub.2) to
provide imine intermediate E1. Imine intermediate E1 was treated
with TMSCN to provide furopyridine (I-C).
##STR00043##
[0177] Scheme 4A depicts a synthesis of furopyridine derivative
(I-L). To compound 4-1 was added potassium tert-butoxide.
Methoxymethyl chloride was then added to form MOM protected
compound 4-2. LDA was then added to the solution of compound 4-2.
N-formylpiperidine was then added to this solution resulting in the
formation of compound 4-3. Aldehyde 4-3 was then deprotected using
an acid to compound 4-4. In one embodiment, the deprotection was
performed using 3N HCl. Deprotected aldehyde 4-4 was coupled with
an amine (R.sup.5--NH.sub.2) to provide imine intermediate E2. The
corresponding imine intermediate was treated with TMSCN to provide
furopyridine (I-L) via a Strecker reaction followed by
intramolecular cyclization.
##STR00044##
[0178] Scheme 4B depicts a synthesis of furopyridine derivative
4-6. To 3-chloro-5-hydroxypyridine 4-1 was added potassium
tert-butoxide, and then methoxymethyl chloride resulting in the
formation of MOM protected compound 4-2. LDA was added to compound
4-2. N-formylpiperidine was then added resulting in the formation
of compound 4-3. The protected aldehyde 4-3 was deprotected to
afford 3-chloro-5-hydroxy-pyridine-4-carbaldehyde 4-4 using HCl.
The deprotected aldehyde 4-4 was coupled with
3-chloro-4-fluoroaniline to provide imine intermediate 4-5. The
imine intermediate was then treated with TMSCN to provide
furopyridine 4-6.
##STR00045##
[0179] Scheme 5 depicts a synthesis of compound (I-C). NBS or
Ar-iodo succinimide was added to a compound H1 to afford compound
J1. Compound J1 was then reacted with bis(pinacolato)diborane,
potassium or sodium acetate, and Pd(dppf)Cl.sub.1. DCM,
Pd(OAc).sub.2, or Pd.sub.2(dba).sub.3 to provide compound K1.
Compound K1 was then reacted with sodium perborate tetrahydrate to
provide compound B1 which was protected as the MOM ether using a
potassium or sodium tert-butoxide and methoxymethylchloride or
SEMCl. Resultant compound C1 was formylated via reaction with TMEDA
or HMPA, followed by reaction with n-BuLi to provide compound D1.
Corresponding aldehyde D1 was deprotected using an acid to provide
compound A1. Compound A1 was then reacted with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate E1. Corresponding
imine E1 was converted to compound (I-C) using a trialkylsilyl
cyanide such as TMSCN.
##STR00046##
[0180] Scheme 5A depicts the synthesis of compound (I-M). NBS was
reacted with an R.sup.1-substituted pyridine to afford compound M1.
Compound M1, bis(pinacolato)diborane, potassium acetate, and
Pd(dppf)Cl.sub.2. DCM were reacted to provide compound N1. Compound
N1 was reacted with sodium perborate tetrahydrate in water to
provide compound O1 which was then protected as a MOM ether using
potassium tert-butoxide and methoxymethylchloride. The resultant
compound P1 was formylated via its reaction with TMEDA, followed by
addition of n-BuLi to provide compound Q1. Corresponding aldehyde
Q1 was then deprotected using an acid to provide compound R1.
Compound R1 was then reacted with an amine (R.sup.5--NH.sub.2) to
provide imine intermediate S1. The corresponding imine Si was then
converted to compound (I-M) using TMSCN.
##STR00047##
[0181] Scheme 5B provides the synthesis of compound 5-9. NBS was
added to 2-methoxypyridine in MeCN to afford compound 5-2.
5-Bromo-2-methoxypyridine on reaction with bis(pinacolato)diborane
and potassium acetate in the presence of Pd(dppf)Cl.sub.2. DCM
provided
2-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]-dioxaborolan-2-yl)-pyridine
5-3. Compound 5-3 was added to a suspension of sodium perborate
tetrahydrate in water to provide 6-methoxypyridin-3-ol 5-4.
Compound 5-4 was protected as the MOM ether using potassium
tert-butoxide and methoxymethylchloride. Compound 5-5 was then
formylated by adding TMEDA, n-BuLi, and DMF to provide
2-methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde 5-6. Aldehyde
5-6 was deprotected using 3N HCl to provide
5-hydroxy-2-methoxy-pyridine-4-carbaldehyde 5-7. Compound 5-7 was
then treated with 3-chloro-4-fluoroaniline to provide imine
intermediate 5-8. The imine was converted to
N.sup.3-(3-chloro-4-fluoro-phenyl)-5-methoxy-furo[2,3-e]pyridine-2,3-diam-
ine 5-9 using TMSCN.
##STR00048##
[0182] In Scheme 6, DIPEA or TEA was reacted with compound B1. The
addition of methoxymethyl chloride or SEMCl provided the MOM or SEM
protected compound C1. TMEDA was then added to compound C1,
followed by an alkyl lithium reagent such as n-BuLi or sec-BuLi,
and finally followed by DMF to provide MOM protected carbaldehyde
D1. Compound D1 was deprotected using an acid to provide compound
A1. Compound A1 was then treated with an amine (R.sup.5--NH.sub.2)
to provide imine intermediate compound E1. Compound (I-N) was
synthesized by treating corresponding imine E1 with a trialkylsilyl
cyanide such as TMSCN.
##STR00049##
[0183] In Scheme 6A, DIPEA was added to 3-hydroxy-2-substituted
pyridine B2. Methoxymethyl chloride was then added to provide the
MOM protected compound C2. TMEDA was added to compound C2, followed
by n-BuLi, which was followed by DMF to provide MOM protected
carbaldehyde D2. Compound D2 was deprotected using 3N HCl to
provide compound A2. Compound A2 was then reacted with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate E2. Compound
(I-O) was synthesized by reacting imine E2 with TMSCN.
##STR00050##
[0184] In Scheme 6B, DIPEA was added to 3-hydroxy-2-methylpyridine
6-1. Methoxymethyl chloride was then added to provide MOM protected
compound 6-2. TMEDA was then added, n-BuLi (2.17 M in hexane) was
added, and DMF was then added to provide the MOM protected
carbaldehyde 6-3 which was deprotected using 3N HCl to provide
3-hydroxy-2-methylpyridine-4-carbaldehyde 6-4. Compound 6-4 was
then treated with 3-chloro-4-fluoroaniline, resulting in the
formation of imine intermediate 6-5.
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-methyl-furo[2,3-c]pyridine-2,3-diami-
ne 6-6 was synthesized by treating imine 6-5 with TMSCN.
##STR00051##
[0185] Scheme 7 provides the synthesis of compound (I-C).
Specifically, compound J1, bis(pinacolato)diborane, potassium or
sodium acetate, and Pd(dppf)Cl.sub.2. DCM, Pd(OAc).sub.2,
Pd(dba).sub.3, or Pd.sub.2(dba).sub.3 were reacted to provide
(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine K1.
Compound K1 was reacted with sodium perborate tetrahydrate to
provide pyridinol or phenol compound B1. Compound B1 was combined
with potassium or sodium alkoxide such as potassium tert-butoxide,
methoxymethyl chloride, and TMEDA or HMPA to provide compound C1.
Compound C1 was then reacted with an alkyl lithium reagent such as
n-BuLi or s-BuLi, followed by DMF or n-formylpiperidine to provide
compound D1. Compound D1 was then deprotected using an acid to
provide compound A1. Compound A1 was then treated with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate E1, which was
treated with a trialkylsilyl cyanide such as TMSCN to provide
compound (I-C).
##STR00052##
[0186] Scheme 7A provides the synthesis of compound (I-P).
Specifically, R.sup.1-substituted 3-bromo-pyridine U2,
bis(pinacolato)diborane, potassium acetate, and Pd(dppf)Cl.sub.2.
DCM were reacted to provide
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine V2.
Compound V2 was reacted with sodium perborate tetrahydrate in water
to provide R.sup.1-substituted- pyridin-3-ol W2. Compound W2 was
stepwise combined with potassium tert-butoxide, methoxymethyl
chloride, and TMEDA to provide compound X2. Compound X2 was then
reacted with n-BuLi, followed by DMF to provide compound Y2.
Compound Y2 was then deprotected using an acid to provide compound
Z2. Compound Z2 was then treated with an amine (R.sup.5--NH.sub.2)
to provide imine intermediate AA2, which was treated with TMSCN to
provide compound (I-P).
##STR00053##
[0187] Scheme 7B provides the synthesis of compound 7-8.
3-Bromo-5-methoxypyridine 7-1, bis(pinacolato)diborane, potassium
acetate, and Pd(dppf)Cl.sub.2. DCM were reacted to provide
3-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
7-2. Compound 7-2 was reacted with a suspension of sodium perborate
tetrahydrate in water to provide 5-methoxy-pyridin-3-ol 7-3.
Compound 7-4 was prepared by reacting compound 7-3, potassium
tert-butoxide, and methoxymethyl chloride. Compound 7-4 was
lithiated using n-BuLi and TMEDA and the resultant lithiated
species was quenched with DMF to yield
3-methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde 7-5. Compound
7-5 was deprotected using 3N HCl to provide
3-hydroxy-5-methoxy-pyridine-4-carbaldehyde 7-6. Compound 7-6 was
then treated with 3-chloro-4-fluoroaniline to form imine
intermediate 7-7 which was treated with TMSCN to provide
N.sup.3-(3-chloro-4-fluoro-phenyl)-4-methoxy-furo[2,3-c]pyridine-2,3-diam-
ine 7-8.
##STR00054##
[0188] Scheme 8 describes the synthesis of compound (I-Q).
Specifically, an acid was bubbled through compound BB1 to provide
compound CC1 in acetone solution. In one embodiment, the acid was
dry HCl. Thionyl chloride or oxalyl chloride in the presence of a
catalytic amount of DMF was then added to compound CC1 to provide
compound DD1. Compound DD1 was hydrogenated with hydrogen using a
catalyst, such as Pd/C and sodium or potassium acetate to provide
compound EE1. The alcoholic group of compound EE1 was oxidized to
provide aldehyde FF1. In one embodiment, the oxidation was
performed using manganese dioxide or pyridinium chloro chromate.
Compound FF1 was then coupled with an amine (R.sup.5--NH.sub.2) to
provide an imine intermediate which underwent in-situ Strecker
reaction followed by intramolecular cyclization with TMSCN in the
presence of acetic acid to afford compound (I-Q).
##STR00055##
[0189] Scheme 8A describes the synthesis of compound (I-R) via
treatment of an acetone solution of compound BB2 with an anhydrous
acid such as HCl gas or sulfuric acid, to provide compound CC2.
Thionyl chloride was then added to compound CC2 to provide compound
DD2. Compound DD2 was hydrogenated with hydrogen gas using a
catalyst, such as Pd/C and sodium acetate, to provide compound EE2.
The alcoholic group of compound EE2 was oxidized with a reagent
such as manganese dioxide to provide aldehyde FF2. Compound FF2 was
then coupled with an amine (R.sup.5--NH.sub.2) to provide an imine
intermediate which undergoes in-situ Strecker reaction followed by
intramolecular cyclization with TMSCN in the presence of acetic
acid to afford compound (I-R).
##STR00056##
[0190] Scheme 8B describes the synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-4,7
dimethylfuro[2,3-c]pyridine-2,3-diamine 8-6. Dry HCl was bubbled
through a suspension of pyridoxine hydrochloride 8-1 in acetone to
afford 2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridin-5-yl)methanol
hydrochloride 8-2. Thionyl chloride was then added to the solution
of compound 8-2 to afford
5-(chloromethyl)-2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridine
hydrochloride 8-3. Compound 8-3 was hydrogenated with Pd/C and
sodium acetate under hydrogen, resulting in the formation of
4-(hydroxymethyl)-2,5-dimethylpyridin-3-ol 8-4. The alcoholic group
of compound 8-4 was oxidized with manganese dioxide to give
aldehyde 8-5. 3-Hydroxy-2,5-dimethylisonicotinaldehyde 8-5 was
coupled with 3-chloro-4-fluoroaniline resulting in the formation of
imine intermediate which undergoes in situ Strecker reaction
followed by intramolecular cyclization with TMSCN to afford
N.sup.3-(3-chloro-4-fluorophenyl)-4,7-dimethylfuro[2,3-c]pyridine-2,3-dia-
mine 8-6.
##STR00057##
[0191] The synthesis of compound (I-S) is provided in Scheme 9. A
strong base such as sodium or potassium alkoxide was added to
compound GG1. In one embodiment, the strong base was potassium
tert-butoxide. Methoxymethyl chloride was then added dropwise
resulting in the formation of MOM protected compound HH1. To a
stirred solution of compound HH1 was added an R.sup.4-substituted
alkylating agent and a catalyst to provide compound II1. In one
embodiment, the alkylating agent was triethylborate. In another
embodiment, the catalyst was Pd(PPh.sub.3).sub.4. TMEDA or HMPA was
then added to stirred solution of compound II1. n-BuLi, s-BuLi,
LDA, or LTMP was then added, followed by DMF or N-formylpiperidine
to afford compound JJ1. Compound JJ1 was deprotected to provide
compound KK1. In one embodiment, this reaction was performed using
an acid. In another embodiment, the acid was HCl. Compound KK1 was
then treated with an amine (R.sup.5--NH.sub.2) to provide imine
intermediate LL1. Compound LL1 was then converted to compound (I-S)
using a trialkylsilyl cyanide such as TMSCN.
##STR00058##
[0192] The synthesis of compound (I-O) is provided in Scheme 9A.
Potassium tert-butoxide was added to a solution of compound GG2.
Methoxymethyl chloride was then added resulting in the formation of
MOM protected compound HH.sub.2. To compound HH2 was added an
R.sup.4-substituted alkylating agent, such as triethylborate,
K.sub.2CO.sub.3 and Pd(PPh.sub.3).sub.4 to provide compound C2.
TMEDA was then added to compound C2. n-BuLi was then added,
followed by DMF to afford compound D2. Compound D2 was deprotected
to provide compound I2. Compound I2 was then treated with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate E2. Compound E2
was then converted to compound (I-O) using TMSCN.
##STR00059##
[0193] The synthesis of compound 9-7 is depicted in Scheme 9B.
Potassium tert-butoxide was added to compound 9-1. Methoxymethyl
chloride was then added, resulting in the formation of MOM
protected compound 9-2. To a solution of
2-chloro-3-methoxymethoxy-pyridine 9-2 was added triethylborate,
K.sub.2CO.sub.3 and Pd(PPh.sub.3).sub.4 to afford
2-ethyl-3-methoxymethoxypyridine 9-3. TMEDA was added, followed by
n-BuLi, and then DMF to afford 2-ethyl-3-methoxy
methoxypyridine-4-carbaldehyde 9-4. Compound 9-4 was deprotected
using 3N HCl to provide 2-ethyl-3-hydroxypyridine-4-carbaldehyde
9-5. Compound 9-5 was treated with 3-chloro-4-fluoroaniline to
provide imine intermediate 9-6 which is in-turn converted to
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-ethyl-furo[2,3-c]pyridine-2,3-diamin-
e 9-7 in the presence of TMSCN.
##STR00060##
[0194] Scheme 10 describes another synthesis of compound (I-S).
Compound HH1 was prepared by adding a base and methoxymethyl
chloride to compound GG1. In one embodiment, the base was any of
potassium tert-butoxide, NaH, or potassium carbonate. A catalyst
and an R.sup.4-substituted alkylating agent were then added to
provide compound II1. In one embodiment, the alkylating agent was
propyl magnesium chloride. In other embodiments, the alkylating
agents are any of methyl magnesium bromide, ethyl magnesium bromide
or aryl magnesium bromide such as phenyl magnesium bromide. In
another embodiment, the catalyst was Ni(dppp)Cl.sub.2. TMEDA or
LTMP was then added to compound II1. n-BuLi or s-BuLi was next
added to the reaction mixture. To the lithiated species, DMF was
added to provide an aldehyde compound JJ1. Aldehyde compound JJ1
was deprotected to provide compound KK1. In one embodiment, the
deprotection was performed using an acid. In another embodiment,
the acid was HCl. In another embodiment, the acid was TFA.
Carbaldehyde KK1 was then treated with and an amine
(R.sup.5--NH.sub.2) to provide imine intermediate LL1, which was
converted to compound (I-S) using a trialkylsilyl cyanide such as
TMSCN.
##STR00061##
[0195] Scheme 10A describes the synthesis of (I-T). Compound 9-2
was prepared by adding potassium tert-butoxide and methoxymethyl
chloride to a solution of compound 9-1. To a solution of
2-chloro-3-methoxy methoxypyridine 9-2, Ni(dppp)Cl.sub.2 and propyl
magnesium chloride (2M soln) were added to provide
3-methoxymethoxy-2-propylpyridine 10-3. TMEDA and n-BuLi was added
to compound 10-3. To the resulting lithiated species, DMF was added
to provide 3-methoxymethoxy-2-propylpyridine-4-carbaldehyde 10-4.
Aldehyde 10-4 was deprotected to provide
3-hydroxy-2-propylpyridine-4-carbaldehyde 10-5 using 3N HCl.
Carbaldehyde 10-5 was then treated with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate LL2. The compound
LL2 was converted to compound (I-T) using TMSCN.
##STR00062##
[0196] Scheme 10B describes the synthesis of
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-propyl-furo[2,3-c]pyridine-2,3-diami-
ne 10-7. Compound 9-2 was prepared by adding potassium
tert-butoxide and methoxymethyl chloride to a solution of compound
9-1. To a solution of 2-chloro-3-methoxy methoxypyridine 9-2,
Ni(dppp)Cl.sub.2 and propyl magnesium chloride (2M soln) were added
to provide 3-methoxymethoxy-2-propylpyridine 10-3. TMEDA was added
to compound 10-3 and then n-BuLi added. To the lithiated species,
DMF was added to provide
3-methoxymethoxy-2-propylpyridine-4-carbaldehyde 10-4. Aldehyde
10-4 was deprotected to provide
3-hydroxy-2-propylpyridine-4-carbaldehyde 10-5 using 3N HCl.
Carbaldehyde 10-5 was then treated with 3-chloro-4-fluoroaniline to
provide imine intermediate 10-6. 4-
{[3-chloro-4-fluoro-phenylimino]-methyl}-2-propyl-pyridin-3-ol 10-6
was converted to
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-propyl-furo[2,3-c]pyridine-2,3-diami-
ne 10-7 using TM S CN.
##STR00063##
[0197] The synthesis of compound (I-U) is described in Scheme 11. A
base was added into a solution of compound SS1. In one embodiment,
the base was K.sub.2CO.sub.3. Iodine in methanol was then added to
compound SS1 to afford compound TT1. To a stirred solution of
compound TT1 was added methoxymethyl chloride or SEMC1, followed by
an organic base such as DIPEA to yield compound UU1. Compound UU1
was then treated with R.sup.2-substituted alkoxide or aryloxide in
the presence of CuBr to provide the compound VV1 via replacement of
the I-substituent. In one embodiment, the R.sup.2-substituted
alkoxide was sodium methoxide. TMEDA or HMPA was added to compound
VV1 and n-BuLi or s-BuLi was added to provide the lithiated
species. DMF was then added to provide carbaldehyde WW1, which was
deprotected to provide compound XX1 using an acid. In one
embodiment, the acid was HCl. Compound XX1 was reacted with an
amine (R.sup.5--NH.sub.2) to provide imine intermediate YY1.
Compound (I-U) was synthesized by reacting a trialkylsilyl cyanide
such as TMSCN with compound YY1.
##STR00064##
[0198] The synthesis of compound (I-V) is described in Scheme 11A.
K.sub.2CO.sub.3 was added to compound B2. Iodine in methanol was
then added to afford compound TT2. To a solution of compound TT2,
methoxymethyl chloride was added followed by DIPEA to yield
compound UU2. A solution of compound UU2 was added to freshly
prepared sodium methoxide, followed by CuBr to yield compound VV2.
TMEDA was added to compound VV2 and n-BuLi was added. DMF was then
added to provide carbaldehyde WW2, which was deprotected to provide
XX2 by using HCl. Compound XX2 was reacted with an amine
(R.sup.5--NH.sub.2) to provide imine intermediate YY2. Compound
(I-V) was synthesized by adding TMSCN to the solution of compound
YY2.
##STR00065##
[0199] The synthesis of
N.sup.3-(3-chloro-4-fluoro-phenyl)-5-methoxy-7-methyl
-furo[2,3-c]pyridine-2,3-diamine 11-8 is described in Scheme 11B.
K.sub.2CO.sub.3 was added to 2-methyl-pyridine-3-ol 11-1. Iodine in
methanol was then added to afford 6-iodo-2-methylpyridine-3-ol
11-2. To compound 11-2, methoxymethyl chloride was added followed
by DIPEA to yield 6-iodo-3-methoxymethoxy-2-methyl pyridine 11-3.
Compound 11-3 was added to freshly prepared sodium methoxide
followed by CuBr to yield compound 11-4. TMEDA was added to
6-methoxy-3-methoxy methoxy-2-methyl pyridine 11-4 and n-BuLi was
added, followed by DMF to synthesize carbaldehyde 11-5. 6-M
ethoxy-3-methoxymethoxy-2-methylpyridine-4-carbaldehyde 11-5 was
then deprotected to provide
3-hydroxy-6-methoxy-2-methyl-pyridine-4-carbaldehyde 11-6. Compound
11-6 was then reacted with 4-fluoro-3-chloroaniline to give imine
intermediate 11-7. The product 11-8 was synthesized by adding TMSCN
to the solution of compound 11-7.
##STR00066##
[0200] Scheme 12 depicts the synthesis of compound (I-W-1).
Compound ZZ12-1 was treated with a base like n-BuLi, LDA, etc.
followed by addition of an awl or heteroarylnitrile. In another
embodiment an acid halide or Weinreb amide can also be used. The
mixture was then quenched with ammonium chloride and pH adjusted to
about 2 to about 4 using an acid to provide compound ZZ12-2. In one
embodiment, the pH was adjusted to 3. In another embodiment, the
acid was 6N HCl. See, Chubb et al., J. Chem. Soc., Perkin. Trans.,
1853-1854 (2001), which is hereby incorporated by reference.
[0201] Compound ZZ12-3 was obtained by treating compound ZZ12-2
with ammonium hydroxide in an autoclave at 100-140.degree. C.
Protection of the OH group in compound ZZ12-3 with MOM or SEM to
provide compound ZZ12-4 was performed with a potassium or sodium
alkoxide such as tert-BuOK and methoxymethyl chloride or SEMC1.
Treatment of protected compound ZZ12-4 with TMP or TMEDA and n-BuLi
followed by addition of DMF generated protected aldehyde ZZ12-5.
Deprotection was done with an acid to provide compound ZZ12-6. In
one embodiment, the acid is hydrochloric acid. Treatment of
aldehyde ZZ12-6 with an amine (R.sup.5--NH.sub.2) provided imine
ZZ1 which was further reacted with TMSCN to provide target compound
(I-W-1).
##STR00067##
[0202] Scheme 12 depicts the synthesis of compound (I-W). Furan
12-1 was treated with n-BuLi, followed by addition of
3-cyanopyridine. The mixture was then quenched with ammonium
chloride and pH adjusted to about 2 to about 4 using an acid to
provide compound 12-2. In one embodiment, the pH was adjusted to 3.
In another embodiment, the acid was 6N HCl. See, Chubb et al., J.
Chem. Soc., Perkin. Trans., 1853-1854 (2001), which is hereby
incorporated by reference. Compound 12-3 was obtained by treating
compound 12-2 with ammonium hydroxide in an autoclave at
140.degree. C. Protection of the OH group in compound 12-3 with MOM
or SEM to provide compound 12-4 was performed with a potassium or
sodium alkoxide such as tert-BuOK and methoxymethyl chloride or
SEMC1. Treatment of protected compound 12-4 with TMP or TMEDA and
n-BuLi followed by addition of DMF generated protected aldehyde
12-5. Deprotection was done with an acid to provide compound 12-6.
In one embodiment, the acid is hydrochloric acid. Treatment of
aldehyde 12-6 with an amine (R.sup.5--NH.sub.2) provided imine
intermediate ZZ1 which was further reacted with TMSCN to provide
target compound (I-W).
[0203] Scheme 12B depicts the synthesis of compound 12-8. Furan
12-1 was treated with n-BuLi, followed by addition of
3-cyanopyridine. The mixture was then quenched with ammonium
chloride and pH adjusted to 3 using 6N HCl to provide compound
12-2. Compound 12-3 was obtained by treating compound 12-2 with
ammonium hydroxide in an autoclave at 140.degree. C. Protection of
the c OH group in compound 12-3 with MOM to provide compound 12-4
was performed with tert-BuOK and methoxymethyl chloride. Treatment
of MOM protected compound 12-4 with TMP and n-BuLi followed by
addition of DMF generated the MOM protected aldehyde 12-5.
Deprotection was done with 3N HCl to provide compound 12-6.
Treatment of aldehyde 12-6 with 4-fluoro-3-chlorophenylamine
provided imine 12-7 which was further reacted with TMSCN to provide
the target compound 12-8.
##STR00068##
[0204] Scheme 13 shows the preparation of cyano-substituted
compound (I-X-1). Compound AAA1-1 was treated with a base, followed
by addition of methoxymethylchloride to provide the MOM protected
product BBB1-1. In one embodiment, the base was sodium or potassium
tert-butoxide. Compound BBB1-1 was treated with TMP or LDA and
n-BuLi, followed by DMF or N-formylpiperidine to provide compound
CCC1-1. Compound CCC1-1 was deprotected to DDD1-1, and then
condensed with amine (R.sup.5--NH.sub.2) to provide compound
EEE1-1. EEE1-1 was further reacted with TMSCN under Strecker
reaction conditions to provide compound (I-X-1).
##STR00069##
[0205] Scheme 13A1 shows the preparation of cyano-substituted
compound (I-X). Compound AAA1 was treated with a base, followed by
addition of methoxymethylchloride to provide the MOM protected
product BBB1. In one embodiment, the base was sodium or potassium
tert-butoxide. Compound BBB1 was treated with TMP or LDA and
n-BuLi, followed by DMF or N-formylpiperidine to provide compound
CCC1. Compound CCC1 was deprotected and condensed with an amine
(R.sup.5--NH.sub.2) provide compound EEE 1 which was further
reacted with TMSCN under Strecker reaction condition to provide
compound (I-X).
##STR00070##
[0206] Scheme 13A2 shows the preparation of cyano-substituted
compound (I-Y). Compound AAA2 was treated with potassium
tert-butoxide, followed by addition of methoxymethylchloride to
provide the MOM protected product BBB2. Compound BBB2 was treated
with TMP and n-BuLi, followed by DMF to provide compound CCC2.
Compound CCC2 was deprotected and condensed with an amine
(R.sup.5--NH.sub.2) to provide compound EEE2 which was further
reacted with TMSCN under Strecker reaction condition to provide
compound (I-Y).
##STR00071##
[0207] Scheme 13B shows the preparation of cyano-substituted
compound 13-6. 2-Cyano-3-hydroxypyridine 13-1 was treated with
potassium tert-butoxide, followed by addition of
methoxymethylchloride to provide the MOM protected product 13-2.
Compound 13-2 was treated with TMP and n-BuLi in THF followed by
DMF to provide formylated product 13-3. Compound 13-3 was
deprotected and condensed with 3-chloro-4-fluoroaniline to provide
compound 13-5 which was further reacted with TMSCN under Strecker
reaction condition to provide compound 13-6.
##STR00072##
[0208] Scheme 14 provides the preparation of compound (I-U).
Specifically, compound EEEE1-1 prepared as described above was
reacted with an R.sup.4-substituted boronic acid and a Pd catalyst.
In one embodiment, the catalyst was Pd(PPh.sub.3).sub.4. Product
SS1 was iodinated to provide compound TT1. In one embodiment, the
iodination was performed using with iodine. Iodo compound TT1 was
protected by MOMCl and then R.sup.2-substituted in presence of a Pd
catalyst or [1,3-bis(diphenylphosphino)propane]dichloronickel (II)
to provide compound VV1. In one embodiment, the catalyst was
[1,3-bis(diphenylphosphino)propane]dichloronickel (II). In another
embodiment, the R.sup.2-substitution was performed using an
alkylated borate such as triethylborate. Compound VV1 was lithiated
using n-BuLi and formulated using DMF to provide compound XX1. In
one embodiment, the formylation was performed in TMEDA using DMF as
the formylating agent. The formylated product XX1 was converted to
compound (I-U) via intermediate YY1 as described above.
##STR00073##
[0209] Scheme 14A1 provides the preparation of compound (I-Z).
Specifically, compound EEEE1, prepared as described above was
reacted with an R.sup.4-substituted boronic acid and a Pd catalyst.
In one embodiment, the catalyst was Pd(PPh.sub.3).sub.4. Product
WWW1 was iodinated to provide compound XXX1. In one embodiment, the
iodination was performed using with iodine. Iodo compound XXX1 was
protected as a MOM ether and then R.sup.2-substituted in presence
of a Pd catalyst or
[1,3-bis(diphenylphosphino)propane]dichloronickel (II) to provide
compound ZZZ1. In one embodiment, the catalyst was
[1,3-bis(diphenylphosphino)propane]dichloronickel (II). In another
embodiment, the R.sup.2 substitution was performed using an
alkylated borate such as triethylborate. Compound ZZZ1 was
lithiated using n-BuLi and formulated using DMF to provide compound
BBBB1. In one embodiment, the formylation was performed in TMEDA
using DMF as the formulation agent. The formylated product BBBB1
was converted to compound (I-Z) via intermediate YY2 as described
above.
##STR00074##
[0210] Scheme 14A2 provides the preparation of compound (I-V).
Specifically, 2-iodo-3-hydroxypyridine 14-2 prepared as described
above was reacted with an R.sup.4-substituted boronic acid and a
catalyst. In one embodiment, the catalyst was Pd(PPh.sub.3).sub.4.
Product B2 was iodinated with iodine to provide compound TT2.
Compound TT2 was MOM protected via a coupling reaction and then
R.sup.2-substituted in presence of a catalyst to provide compound
VV2. In one embodiment, the catalyst was
[1,3-bis(diphenylphosphino)propane]dichloronickel (II). In another
embodiment, the R.sup.2-substitution was performed using
triethylborate. Formylation of compound VV2 was done by reaction
with n-BuLi and DMF to provide compound WW2. In one embodiment, the
formylation was performed in TMEDA using N-formylpiperidide. The
formylated product WW2 was converted to compound (I-V) via
intermediate YY2 as described above.
##STR00075## ##STR00076##
[0211] Scheme 14B provides the preparation of compound 14-10.
Specifically, 2-iodo-3-hydroxypyridine 14-2 prepared as described
above was refluxed with phenylboronic acid and sodium bicarbonate
solution in presence of Pd(PPh.sub.3).sub.4. Product 14-3 was
iodinated with iodine in presence of sodium bicarbonate to provide
compound 14-4. Iodo compound 14-4 was MOM protected to give 14-5
which was converted to 14-6 via a coupling reaction with
triethylborate in presence of
[1,3-bis(diphenylphosphino)propane]dichloronickel(II) to provide
compound 14-6. Reaction of compound 14-6 with n-BuLi in TMEDA
provided the lithiated species which was formylated using DMF. The
formylated product 14-8 was converted to compound 14-10 via
intermediate 14-9 as described herein.
##STR00077##
[0212] Scheme 15 describes the preparation of compound (I-HH).
Compound JJJ1 was treated with bromoacetaldehyde diethyl acetal in
the presence of potassium carbonate in DMF to afford compound KKK1.
Compound KKK1 was refluxed in the presence of PPA or a Lewis acid
such as ZnCl.sub.2, AlCl.sub.3, or AlBr.sub.3 to give compound
LLL1. In one embodiment, this reaction was performed in a high
boiling aromatic solvent such as chlorobenzene, toluene, xylenes,
or diphenylether. Compound LLL1 was then R.sup.2-substituted via a
Suzuki-Miyaura cross coupling reaction in the presence of a
catalyst such as Pd(PPh.sub.3).sub.4, Pd(OAc).sub.2, or
PdCl.sub.2(PPh.sub.3).sub.2 in the presence of potassium carbonate
in refluxing dioxane to afford compound FFF1. Compound FFF1 was
brominated to give compound GGG1. In one embodiment, the
bromination is performed using NBS in the presence of chloroform
and acetic acid. Compound GGG1 was then nitrated using fuming
nitric acid and acetic acid or fuming nitric acid and TFA or fuming
nitric acid and sulfuric acid to afford compound HHH1. Compound
HHH1 was NHR.sup.5-substituted using an amine (R.sup.5--NH.sub.2)
to form compound III1. Catalytic hydrogenation was then done on
compound III1 using Pd/C (10%) under hydrogen gas in methanol,
ethanol, isopropanol, or ethyl acetate to afford compound
(I-HH).
##STR00078##
[0213] Scheme 15A describes the preparation of compound (I-II).
Commercially available 4-bromothiophenol 15-1 was treated with
bromoacetaldehyde diethyl acetal in the presence of potassium
carbonate in DMF to afford compound
15-2.1-Bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene 15-2 was
refluxed in the presence of PPA or a Lewis acid such as ZnCl.sub.2,
AlCl.sub.3, or AlBr.sub.3 to give 5-bromo-1-benzothiophene 15-3. In
one embodiment, this reaction was performed in a high boiling
aromatic solvent such as chlorobenzene, toluene, xylene, or
diphenylether. Compound 15-3 was then R.sup.2-substituted via a
Suzuki-Miyaura cross coupling reaction in the presence of a
catalyst such as Pd(PPh.sub.3).sub.4, Pd(OAc).sub.2, or
PdCl.sub.2(PPh.sub.3).sub.2 in the presence of potassium carbonate
in refluxing dioxane to afford compound FFF2. Compound FFF2 was
brominated to give compound GGG2. In one embodiment, the
bromination is performed using NBS in the presence of chloroform
and acetic acid. Compound GGG2 was then nitrated using fuming
nitric acid and acetic acid or fuming nitric acid and TFA or fuming
nitric acid and sulfuric acid to afford compound HHH.sub.2.
Compound HHH.sub.2 was NHR.sup.5-substituted using an amine
(R.sup.5--NH.sub.2) to give compound III2. Catalytic hydrogenation
was then done on compound III2 using Pd/C (10%) under hydrogen gas
in methanol, ethanol, isopropanol or ethyl acetate to afford
compound (I-II).
##STR00079##
[0214] Scheme 15B describes the preparation of
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenyl-1-benzothiophene-2,3-diamine
15-8. 4-Bromothiophenol 15-1 was treated with bromoacetaldehyde
diethyl acetal in the presence of potassium carbonate in DMF to
afford 15-2.1-Bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene 15-2 was
refluxed in the presence of PPA or a Lewis acid such as ZnCl.sub.2,
AlCl.sub.3, or AlBr.sub.3 to give 5-bromo-1-benzothiophene 15-3. In
one embodiment, this reaction was performed in a high boiling
aromatic solvent such as chlorobenzene, toluene, xylene, or
diphenylether. Compound 15-3 was then subjected to Suzuki-Miyaura
cross coupling reaction with phenylboronic acid in the presence of
a catalyst such as Pd(PPh.sub.3).sub.4, Pd(OAc).sub.2, or
PdCl.sub.2(PPh.sub.3).sub.2 in the presence of potassium carbonate
in refluxing dioxane to afford 5-phenyl-1-benzothiophene 15-4.
Compound 15-4 was brominated to give
3-bromo-5-phenyl-1-benzothiophene 15-5. In one embodiment, the
bromination is performed using NBS in the presence of chloroform or
acetic acid. Compound 15-5 was then nitrated using fuming nitric
acid and acetic acid or fuming nitric acid and TFA or fuming nitric
acid and sulfuric acid to afford
3-bromo-2-nitro-5-phenyl-1-benzothiophene 15-6. Compound 15-6 was
coupled with 3-chloro-4-fluoroaniline in DMF to give
N-(3-chloro-4-fluorophenyl)-2-nitro-5-phenyl-1-benzothiophen-3-amine
15-7. Catalytic hydrogenation was then done on compound 1-7 using
Pd/C (10%) under hydrogen gas in methanol, ethanol, isopropanol or
ethyl acetate to afford
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenyl-1-benzothiophene-2,3-diamine
15-8.
##STR00080##
[0215] Scheme 16 depicts the synthesis of compound (I-JJ-1).
Compound NNN1-1 was chlorinated to give compound OOO1-1. In one
embodiment, the chlorination was performed using hexachloroethane
or N-chlorosuccinimide using n-BuLi, s-BuLi, LTMP, LDA, LTMS, or
LHMDS as the lithiating agent. Compound OOO1-1 was treated with
methyl thioglycolate in the presence of a base such as potassium
carbonate, TEA, NaH, or potassium t-butoxide to afford compound
PPP1-1. Compound PPP1-1 underwent diazotization with sodium nitrite
and hydrobromic acid resulting in the formation of diazonium salt
which was reacted with copper bromide to give compound QQQ1-1.
Compound QQQ1-1 was subjected to a Buchwald-Hertwig amination
reaction with an amine (R.sup.5--NH.sub.2) in the presence of
catalyst, such as Pd.sub.2(dba).sub.3 or Pd(dba).sub.2, and BINAP
or XPhos to afford compound JJJ1-1. Compound JJJ1-1 then underwent
Boc protection with di-tent-butyl dicarbonate in the presence of a
base such as TEA, potassium carbonate, DIPEA, sodium carbonate,
NaH, sodium t-butoxide, or potassium t-butoxide followed by
hydrolysis to give compound LLL1-1. Compound LLL1-1 was treated
with an azide source such as DPPA and DIPEA under Curtius
rearrangement condition to afford compound MMM1-1. Boc deprotection
was then performed on compound MMM1-1 using hydrochloric acid or
TFA in a solvent such as dioxane, THF, or DCM to afford compound
(I-JJ-1).
##STR00081##
[0216] Scheme 16A1 depicts the synthesis of compound (I-JJ).
Compound NNN1 was chlorinated to give compound OOO1. In one
embodiment, the chlorination was performed using hexachloroethane
or N-chlorosuccinimide using n-BuLi, s-BuLi, LTMP, LDA, LTMS, or
LHMDS as the lithiating agent. Compound OOO1 was treated with
methyl thioglycolate in the presence of a base such as potassium
carbonate, TEA, NaH, or potassium t-butoxide to afford compound
PPP1. Compound PPP1 underwent diazotization with sodium nitrite and
hydrobromic acid resulting in the formation of diazonium salt which
was reacted with copper bromide to give compound QQQ1. Compound
QQQ1 was subjected to a Buchwald-Hertwig amination reaction with an
amine (R.sup.5--NH.sub.2) in the presence of catalyst, such as
Pd.sub.2(dba).sub.3 or Pd(dba).sub.2, and BINAP or XPhos to afford
compound JJJ1. Compound JJJ1 then underwent Boc protection with
di-tert-butyl dicarbonate in the presence of a base such as TEA,
potassium carbonate, DIPEA, sodium carbonate, NaH, sodium
t-butoxide, or potassium t-butoxide followed by hydrolysis to give
compound LLL1. Compound LLL1 was treated with an azide source such
as DPPA and DIPEA under Curtius rearrangement condition to afford
compound MMM1. Boc dcprotcction was then performed on compound MMM1
using hydrochloric acid or TFA in a solvent such as dioxane, THF,
or DCM to afford compound (I-JJ).
##STR00082##
[0217] Scheme 16A2 depicts the synthesis of compound (1-EE).
4-Cyanopyridine 16-1 was chlorinated to give
3-chloropyridine-4-carbonitrile 16-2. In one embodiment, the
chlorination was performed using hexachloroethane or
N-chlorosuccinimide in the presence of a lithiating reagent such as
n-BuLi, s-BuLi, LTMP, LDA, LTMS, or LHMDS as the lithiating agent.
Compound 16-2 was treated with methyl thioglycolate in the presence
of potassium carbonate, TEA, NaH, or potassium t-butoxide to afford
methyl 3-bromothieno[2,3-c]pyridine-2-carboxylate 16-3. Compound
16-3 underwent diazotization with sodium nitrite and hydrobromic
acid resulting in the formation of the diazonium salt which was
reacted with copper bromide to give compound 16-4.
Methyl-3-aminothieno [2,3-c]pyridine-2-carboxylate 16-4 was
subjected to a Buchwald-Hertwig amination reaction with an amine
(R.sup.5--NH.sub.2) in the presence of catalyst such as
Pd.sub.2(dba).sub.3 or Pd(dba).sub.2 and BINAP or XPhos to afford
compound JJJ2. Compound JJJ2 then underwent Boc protection with
di-tent-butyl dicarbonate in the presence of a base such as TEA,
potassium carbonate, DIPEA, sodium carbonate, NaH, sodium
t-butoxide, or potassium t-butoxide followed by hydrolysis to give
compound LLL2. Compound LLL2 was treated with an azide source such
as DPPA and DIPEA under Curtius rearrangement condition to afford
compound MMM2. Boc deprotection was then performed on compound MMM2
using hydrochloric acid or TFA in a solvent such as dioxane, THF,
or DCM to afford compound (I-EE).
##STR00083##
[0218] Scheme 16B depicts the synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)thieno[2,3-c]pyridine-2,3-diamine
hydrochloride 16-9. 4-Cyanopyridine 16-1 was chlorinated to give
3-chloropyridine-4-carbonitrile 16-2. In one embodiment, the
chlorination was performed using hexachloroethane or
N-chlorosuccinimide in the presence of a lithiating reagent such as
n-BuLi, s-BuLi, LTMP, LDA, LTMS, or LHMDS as the lithiating agent.
Compound 16-2 was treated with methyl thioglycolate in the presence
of potassium carbonate, TEA, NaH, or potassium t-butoxide to afford
methyl 3-aminothieno [2,3-c]pyridine-2-carboxylate 16-3. Compound
16-3 underwent diazotization with sodium nitrite and hydrobromic
acid resulting in the formation of the diazonium salt which was
reacted with copper bromide to give compound 16-4.
Mcthyl-3-bromothieno[2,3-c]pyridine-2-carboxylate 16-4 was
subjected to a Buchwald-Hertwig amination reaction with
3-chloro-4-fluoroaniline in the presence of catalyst such as
Pd.sub.2(dba).sub.3 or Pd(dba).sub.2 and BINAP or XPhos to afford
methyl
3-((3-chloro-4-fluorophenyl)amino)thieno[2,3-c]pyridine-2-carboxylate
16-5. Compound 16-5 then underwent Boc protection with
di-tent-butyl dicarbonate in the presence of a base such as TEA,
potassium carbonate, DIPEA, sodium carbonate, NaH, sodium
t-butoxide, or potassium t-butoxide, followed by hydrolysis to give
compound 16-7.
3-((Tert-butoxycarbonyl)(3-chloro-4-fluorophenyl)amino)thieno
[2,3-c]pyridine-2-carboxylic acid 16-7 was treated with an azide
source such as DPPA and DIPEA under Curtius rearrangement condition
to afford
tent-butyl(2-((tert-butoxycarbonyl)amino)thieno[2,3-c]pyridin-3-yl)(3-chl-
oro-4-fluorophenyl)carbamate 16-8. Boc de-protection was then
performed on compound 16-8 using hydrochloric acid or TFA in a
solvent such as dioxane, THF, or DCM to afford
N.sup.3-(3-chloro-4-fluorophenyl)thieno[2,3-c]pyridine-2,3-diamine
hydrochloride 16-9.
##STR00084##
[0219] Scheme 17 describes the synthesis of compound (I-KK).
Compound SSS1 was treated with bromoacetaldehyde diethylacetal in
the presence of a base such as sodium hydride, potassium carbonate,
sodium t-butoxide, potassium t-butoxide, or TEA in a solvent such
as DMF, THF, dioxane, or toluene to afford compound TTT1. Compound
TTT1 was added to a refluxed solution of PPA or a Lewis acid such
as ZnCl.sub.2, AlCl.sub.3, or AlBr.sub.3 and the resulting reaction
mixture refluxed overnight to give a mixture of compounds UUU1 and
VVV1. This mixture which was treated with copper cyanide in the
presence of a catalyst such as Pd(PPh.sub.3).sub.4, Pd(dba).sub.2,
Pd.sub.2(dba).sub.3, Pd(OAc).sub.2, PdCl.sub.2(PPh.sub.3).sub.2 in
DMF, THF, or MeCN to afford a mixture of compounds XXX1 and WWW1.
These compounds were separated using methods known in the art such
as column chromatography or prep-HPLC. Compound WWW1 was brominated
with NBS to give compound FFFF1. In one embodiment, the bromination
was performed using NBS or bromine. Compound FFFF1 then underwent
nitration to afford compound GGGG1. In one embodiment, the
nitration was performed using with potassium nitrate/acetic
anhydride or potassium nitrate/TFA. Compound GGGG1 was coupled with
an amine (R.sup.5--NH.sub.2) to give compound RRR1. Catalytic
hydrogenation was then done on compound RRR1 to afford compound
(I-KK). In one embodiment, this reaction was performed in the
presence of Zn dust/NH.sub.4Cl or Sn/acetic acid in methanol,
ethanol, or butanol.
##STR00085##
[0220] Scheme 17A describes the synthesis of compound (I-LL).
Commercially available 3-bromothiophenol 17-1 was treated with
bromoacetaldehyde diethylacetal in the presence of a base such as
sodium hydride, potassium carbonate, sodium t-butoxide, potassium
t-butoxide, or TEA in a solvent such as DMF, THF, dioxane, or
toluene to afford compound
17-2.1-Bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene 17-2 was added
to a refluxed solution of PPA or a Lewis acid such as ZnCl.sub.2,
AlCl.sub.3, or AlBr.sub.3 and the resulting reaction mixture
refluxed overnight to give a mixture of 6-bromobenzo[b]thiophene
17-3 and 4-bromobenzo[b]thiophene 17-4. This mixture which was
treated with copper cyanide in the presence of a catalyst such as
Pd(PPh.sub.3).sub.4, Pd(dba).sub.2, Pd.sub.2(dba).sub.3,
Pd(OAc).sub.2, PdCl.sub.2(PPh.sub.3).sub.2 in a solvent such as
DMF, THF, or MeCN to afford a mixture of
benzo[b]thiophene-6-carbonitrile 17-5 and
benzo[b]thiophene-4-carbonitrile 17-6. These compounds were
separated using methods known in the art such as column
chromatography or prep-HPLC. Compound 17-5 was brominated with NBS
to give 3-bromobenzo[b]thiophene-6-carbonitrile 17-7. In one
embodiment, the bromination was performed using NBS or bromine.
Compound 17-7 then underwent nitration to afford
3-bromo-2-nitrobenzo[b]thiophene-6-carbonitrile 17-8. In one
embodiment, the nitration was performed using with potassium
nitrate/acetic anhydride or potassium nitrate/TFA. Compound 17-8
was coupled with an amine (R.sup.5--NH.sub.2) to give compound
RRR2. Catalytic hydrogenation was then done on compound RRR2 to
afford compound (I-LL). In one embodiment, this reaction was
performed in the presence of Zn dust/NH.sub.4Cl or Sn/acetic acid
in methanol ethanol, or butanol.
##STR00086##
[0221] Scheme 17B describes the synthesis of
2-amino-3-((3-chloro-4-fluorophenyl)-amino)benzo[b]thiophene-6-carbonitri-
le 17-10. 3-Bromothiophenol 17-1 was treated with bromoacetaldehyde
diethylacetal in the presence of a base such as sodium hydride,
potassium carbonate, sodium t-butoxide, potassium t-butoxide, or
TEA in a solvent such as DMF, THF, dioxane, or toluene to afford
compound 17-2.1-Bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene 17-2
was added to a refluxed solution of PPA or a Lewis acid such as
ZnCl.sub.2, AlCl.sub.3, or AlBr.sub.3 and the resulting reaction
mixture refluxed overnight to give a mixture of
6-bromobenzo[b]thiophene 17-3 and 4-bromobenzo[b]thiophene 17-4.
This mixture which was treated with copper cyanide in the presence
of a catalyst such as Pd(PPh.sub.3).sub.4, Pd(dba).sub.2,
Pd.sub.2(dba).sub.3, Pd(OAc).sub.2, PdCl.sub.2(PPh.sub.3).sub.2 in
a solvent such as DMF, THF, or MeCN to afford a mixture of
benzo[b]thiophene-6-carbonitrile 17-5 and
benzo[b]thiophene-4-carbonitrile 17-6. These compounds were
separated using methods known in the art such as column
chromatography or prep-HPLC. Compound 17-5 was brominated with NBS
or bromine to give 3-bromobenzo[b]thiophene-6-carbonitrile 17-7. In
one embodiment, the bromination was performed using NBS or bromine.
Compound 17-7 then underwent nitration to afford
3-bromo-2-nitrobenzo[b]thiophene-6-carbonitrile 17-8. In one
embodiment, the nitration was performed using with potassium
nitrate/acetic anhydride or potassium nitrate/TFA. Compound 17-8
was coupled with 3-chloro-4-fluoroaniline in a solvent such as DMF
to give
3-((3-chloro-4-fluorophenyl)amino)-2-nitrobenzo[b]thiophene-6-carbonitril-
e 17-9. Catalytic hydrogenation was then done on compound 17-9 to
afford
2-amino-3-((3-chloro-4-fluorophenyl)amino)benzo[b]thiophene-6-carbonitril-
e 17-10. In one embodiment, this reaction was performed in the
presence of Zn dust/NH.sub.4Cl or Sn/acetic acid in methanol,
ethanol, or butanol.
##STR00087##
[0222] Scheme 18 depicts the synthesis of compound (I-B). Compound
IIII1 was brominated resulting in the formation of compound JJJJ1.
In one embodiment, the bromination was performed using NBS or
bromine. Compound JJJJ1 underwent nitration to afford nitro
compound KKKK1. In one embodiment, the nitration was performed
using fuming nitric acid and TFA, fuming nitric acid and acetic
acid, or fuming nitric acid and sulfuric acid. Compound KKKK1 was
reacted with R.sup.5--SH to provide compound GGGG1. Compound GGGG1
was hydrogenated resulting in the formation of compound HHHH1. In
one embodiment, the hydrogenation was performed using activated
Pd/C under hydrogen gas or Zn/methanol or Sn/acetic acid. Compound
HHHH1 underwent hydrochloride salt formation to afford compound
(I-B). In one embodiment, salt formation was performed using
hydrochloride gas absorbed in diethyl ether or hydrochloride gas
absorbed in an alcohol such as methanol, ethanol, or isopropanol,
EtOAc, or MTBE.
##STR00088##
[0223] Scheme 18A depicts the synthesis of compound (I-MM).
Commercially available benzothiophene 18-1 was brominated resulting
in the formation of 3-bromo-benzothiophene 18-2. In one embodiment,
the bromination was performed using NBS or bromine. Compound 18-2
underwent nitration to afford nitro compound 18-3. In one
embodiment, the nitration was performed using fuming nitric acid
and TFA, fuming nitric acid and acetic acid, or fuming nitric acid
and sulfuric acid. 3-Bromo-2-nitrobenzothiophene 18-3 was reacted
with R.sup.5-SH to provide compound GGGG2. Compound GGGG2 was
hydrogenated resulting in the formation of compound HHHH2. In one
embodiment, the hydrogenation was performed using activated Pd/C
under hydrogen gas, Zn/methanol or Sn/acetic acid. Compound
HHHH.sub.2 underwent hydrochloride salt formation to afford
compound (I-MM). In one embodiment, salt formation was performed
using hydrochloride gas absorbed in diethyl ether or hydrochloride
gas absorbed in an alcohol such as methanol, ethanol, or
isopropanol, EtOAc, or MTBE.
##STR00089##
[0224] Scheme 18B depicts the synthesis of
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine hydrochloride
18-6. Compound 18-1 was brominated resulting in the formation of
3-bromo-benzothiophene 18-2. In one embodiment, the bromination was
performed using NBS or bromine. Compound 18-2 underwent nitration
to afford nitro compound 18-3. In one embodiment, the nitration was
performed using fuming nitric acid and TFA, fuming nitric acid and
acetic acid, or fuming nitric acid and sulfuric acid.
3-Bromo-2-nitrobenzothiophene 18-3 was reacted with
3-chlorothiophenol in NaOH to provide compound 18-4.
3-[(3-Chlorophenyl)sulfanyl]-2-nitro-1-benzothiophene 18-4 was
hydrogenated resulting in the formation of
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine 18-5. In one
embodiment, the hydrogenation was performed using activated Pd/C
under hydrogen gas, Zn/methanol or Sn/acetic acid. Compound 18-5
underwent hydrochloride salt formation to afford
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine hydrochloride
18-6. In one embodiment, salt formation was performed using
hydrochloride gas absorbed in diethyl ether or hydrochloride gas
absorbed in an alcohol such as methanol, ethanol, or isopropanol,
EtOAc, or MTBE.
##STR00090##
[0225] Scheme 19 describes the synthesis of compound (I-C) or
(I-C-1). Compound A1 or LLLL1 was reacted with an amine
(R.sup.5--NH.sub.2) resulting in the formation of the imine
intermediate which underwent in situ Strecker reaction with a
trialkylsilyl cyanide such as TMSCN or an inorganic cyanide salt
such as NaCN, KCN, or Zn(CN).sub.2 followed by unexpected
intramolecular cyclization in the presence of trimethylsilyl
trifluoromethanesulfonate to afford compound (I-C) or (I-C-1).
##STR00091##
[0226] Scheme 19A describes the synthesis of compound (I-NN).
4-Formyl-3-hydroxybenzonitrile 19-1 was reacted with an amine
(R.sup.5--NH.sub.2) resulting in the formation of the imine
intermediate which underwent in situ Strecker reaction with a
trialkylsilyl cyanide such as TMSCN or an inorganic cyanide salt
such as NaCN, KCN, or Zn(CN).sub.2 followed by unexpected
intramolecular cyclization in the presence of trimethylsilyl
trifluoromethanesulfonate to afford compound (I-NN).
##STR00092##
[0227] Scheme 19B describes the synthesis of
2-amino-3-((3-chloro-4-fluorophenyl)-amino)benzofuran-6-carbonitrile
19-2. 4-Formyl-3-hydroxybenzonitrile 19-1 was reacted with
3-chloro-4-fluoroaniline resulting in the formation of the imine
intermediate which underwent in situ Strecker reaction with a
trialkylsilyl cyanide such as TMSCN or an inorganic cyanide salt
such as NaCN, KCN, or Zn(CN).sub.2 followed by unexpected
intramolecular cyclization in the presence of trimethylsilyl
trifluoromethanesulfonate to afford
2-amino-3-((3-chloro-4-fluorophenyl)amino)benzofuran-6-carbonit-
rile 19-2.
##STR00093##
[0228] Scheme 20 describes the synthesis of compounds (ZZZ1A) and
(ZZZ1B). Compound YYY1 was treated with an alkyl chloroformate in
THF or DCM in the presence of pyridine or TEA to afford the title
compounds. In one embodiment, the alkyl chloroformate was ethyl
chloroformate. In other embodiments, the alkyl chloroformate were
methyl chloroformate or benzyl chloroformate.
##STR00094##
[0229] Scheme 20A describes the synthesis of compounds (ZZZ2A) and
(ZZZ2B). Compound YYY2 was treated with an alkyl chloroformate in
THF or DCM in the presence of pyridine or TEA to afford the title
compounds. In one embodiment, the alkyl chloroformate was ethyl
chloroformate. In other embodiments, the alkyl chloroformate were
methyl chloroformate or benzyl chloroformate.
##STR00095##
[0230] Scheme 20 describes the synthesis of mono-carbamate ethyl
(3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carbamate
20-2 and di-carbamate ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)furo[2,3-c]pyridin-3-y-
l)carbamate 20-3.
N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine
20-1 was treated with ethyl chloroformate in THF in the presence of
pyridine to afford the mono-carbamate 20-2 and di-carbamate 20-3.
One of skill in the art would be able to vary the reaction time and
amounts of reagents utilize to prepare the mono-carbamate,
di-carbamate, or a combination thereof, as needed.
##STR00096##
[0231] Scheme 21 provides the preparation of compound (I-U). The
synthesis began with the iodination of hydroxy compound MMMM1. In
one embodiment iodine and sodium carbonate was used as iodinating
reagent. In another embodiment the iodination was performed in a
mixture of solvents like tetrahydrofuran and water to afforded
compound NNNN1. The resulting compound NNNN1 was forwarded to MOM
protection or SEM protection using MOMCI or SEMCl respectively in
the presence of a base to provide the product OOOO1. The compound
OOOO1 was subjected to a metal catalyzed cross-coupling reaction to
give compound VV1. In one embodiment, the catalyst was
Pd(PPh.sub.3).sub.4. In another embodiment the catalyst was
Pd.sub.2(dba).sub.3. The compound VV1 underwent formylation with
DMF or N-formylpiperidine in the presence of base like n-BuLi,
s-BuLi, LDA, or LTMP at -78.degree. C. to give product WW1. The
compound WW1 was deprotected in presence of a Lewis acid to provide
XX1. In one embodiment the acid was TFA. The compound XX1 was
treated with an amine (R.sup.5NH.sub.2), a cyanide ion source and a
Lewis acid to provide the compound (I-U). In one embodiment the
cyanide ion source was TMSCN. In another embodiment the Lewis acid
was TMSOTf.
##STR00097##
[0232] Scheme 21A provides the preparation of compound (I-V). The
compound hydroxypyridine MMMM2 was iodinated using iodine and
sodium carbonate in a mixture of solvents like tetrahydrofuran and
water to afforded compound NNNN2. The resulting compound NNNN2 was
forwarded to MOM protection or SEM protection using MOMCl or SEMC1
respectively in the presence of a base product OOOO2. The compound
OOOO2 was subjected to a metal catalyzed cross-coupling reaction to
give compound VV2. In another embodiment, the catalyst was
Pd(PPh.sub.3).sub.4. In another embodiment the catalyst was
Pd.sub.2(dba).sub.3. The compound VV2 underwent formylation with
DMF or N-formylpiperidine in the presence of base like n-BuLi,
s-BuLi, LDA, or LTMP at -78.degree. C. to give product WW2. The
compound WW2 was deprotected under acidic conditions to provide
XX2. In one embodiment the acid was TFA. The compound XX2 was
treated with an amine (R.sup.5NH.sub.2), TMSCN and TMSOTf to
provide the compound (I-V) via a sequence of reactions such as
imine formation, Strecker reaction and intramolecular
cyclization.
##STR00098##
[0233] Scheme 21B provides the preparation of compound
N.sup.3-(3-chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine 21-7. The syntheses begin with the iodination of
2-fluoro-5-hydroxypyridine 21-1 using iodine and sodium carbonate
in mixture of solvents of tetrahydrofuran and water to afforded
6-fluoro-3-hydrox-2-iodoypyridine 21-2. The resulting compound 21-2
was forwarded to MOM protection using MOMCl in the presence of
potassium tert-butoxide to give MOM protected product 21-3. The
6-fluoro-2-iodo-3-(methoxymethoxy)pyridine 21-3 was subjected to
Suzuki-Miyaura cross-coupling reaction condition with
4-pyridinylboronic acid in the presence of tripotassium phosphate,
tricyclohexylphosphine and Pd.sub.2(dba).sub.3 in dioxane to afford
6-fluoro-3-(methoxymethoxy)-2,4'-bipyridine 21-4 which was
undergoes formylation with DMF in the presence of n-BuLi at
-78.degree. C. to give formylated product 21-5. The compound 21-5
was treated with TFA-DCM solution to afford MOM-deproted compound
6-fluoro-3-hydroxy-[2,4'-bipyridine]-4-carbaldehyde 21-6. The
compound 21-6 was treated with 3-chloro-4-fluoroaniline, TMSCN,
TMSOTf and DCM in a single pot at room temperature to afford
desired product
N.sup.3-(3-chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine 21-7 as pale yellow solid via a sequence of
reactions such as imine formation, Strecker reaction and
intramolecular cyclization.
##STR00099##
[0234] Scheme 22 provides the preparation of compound (I-QQ). The
synthesis began with the iodination of hydroxy compound SSSS1. In
one embodiment iodine and sodium carbonate was used as iodinating
reagent. In another embodiment the iodination was performed in a
mixture of solvents like tetrahydrofuran and water to afforded
compound TTTT1. The resulting compound TTTT1 was forwarded to MOM
protection or SEM protection using MOMCl or SEMC1 respectively in
the presence of a base to provide the product UUUU1. The compound
UUUU1 underwent formylation with DMF or N-formylpiperidine in the
presence of base like n-BuLi, s-BuLi, LDA, or LTMP at -78.degree.
C. to give product VVVV1. The compound VVVV1 was deprotected in
presence of a Lewis acid to provide WWWW1. In one embodiment the
acid was TFA. The compound WWWW1 was treated with an amine
(R.sup.5--NH.sub.2), a cyanide ion source and a Lewis acid to
provide the compound XXXX1. In one embodiment the cyanide ion
source was TMSCN. In another embodiment the Lewis acid was TMSOTf.
The compound XXXX1 was converted to dicarbamate YYYY1. In one
embodiment ethyl chloroformate in the presence of pyridine in THF
was used for formation of monocarbamate or dicarbamate. The
dicarbamate YYYY1 was converted to the product (I-QQ) under
cross-coupling reaction conditions.
##STR00100##
[0235] Scheme 22A provides the preparation of compound (I-RR). The
synthesis began with the iodination of 3-hydroxypyridine 22-1 which
was treated with iodine in the presence of sodium carbonate in
water to afford 2-iodo-3-hydroxypyridine 22-2. The resulting
compound 22-2 was forwarded to MOM protection using MOMCl in the
presence of potassium tert-butoxide to give MOM protected product
22-3. The 2-iodo-3-(methoxymethoxy)pyridine 22-3 was formylated
with DMF in the presence of LDA in THF at -78.degree. C. to give
2-iodo-3-(methoxymethoxy)-isonicotinaldehyde 22-4 which was
subjected to MOM-de-protection with TFA-DCM to afford
3-hydroxy-2-iodoisonicotinaldehyde 22-5. The compound 22-5 was
treated with an amine (fe-NH.sub.2), a cyanide ion source and a
Lewis acid to provide the compound XXXX2. In one embodiment the
cyanide ion source was TMSCN. In another embodiment the Lewis acid
was TMSOTf. The compound XXXX2 was converted to dicarbamate YYYY2.
In one embodiment ethyl chloroformate in the presence of pyridine
in THF was used for formation of monocarbamate or dicarbamate. The
dicarbamate YYYY2 was converted to the product (I-RR) under
cross-coupling reaction conditions.
##STR00101##
[0236] Scheme 22B described the synthesis of ethyl
(7-((4-carbamoylphenyl)ethynyl)-2-((ethoxycarbonyl)amino)furo[2,3-c]pyrid-
in-3-yl)(3-chloro-4-fluorophenyl)carbamate 22-8. The syntheses
begin with the commercially available 3-hydroxypyridine 14-1 which
was treated with iodine in the presence of sodium carbonate in
water to afford 2-iodo-3-hydroxypyridine 14-2. The resulting
compound 14-2 was forwarded to MOM protection using MOMCl in the
presence of potassium tert-butoxide to give MOM protected product
22-3. The 2-iodo-3-(methoxymethoxy)pyridine 22-3 was formylated
with DMF in the presence of LDA in THF at -78.degree. C. to give
2-iodo-3-(methoxymethoxy)-isonicotinaldehyde 22-4 which was
undergoes MOM-de-protection with TFA-DCM to afford
3-hydroxy-2-iodoisonicotinaldehyde 22-5. The compound 22-5 was
treated in a one-pot with 3-chloro-4-fluoroaniline, TMSCN followed
by TMSOTf in DCM at room temperature to afford
N.sup.3-(3-chloro-4-fluorophenyl)-7-iodofuro[2,3-c]pyridine-2,3-diamine
22-6 via a sequence of reactions such as imine formation, Strecker
reaction and intramolecular cyclization. The compound 22-6
undergoes dicarbamate formation using ethylchloroformate in the
presence of pyridine in THF to afford diethylcarbamate 22-7. The
ethyl (3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-iodo
furo[2,3-c]pyridin-3-yl)carbamate 22-7 was treated under
Sonogashira reaction condition with 4-ethynylbenzamide in the
presence of PdCl.sub.2(PPh.sub.3).sub.2, CuI and triethyl amine to
give desired product ethyl
(7-((4-carbamoylphenypethynyl)-2-((ethoxycarbonyl)amino)furo[2,3-c]pyridi-
n-3-yl)(3-chloro-4-fluorophenyl)carbamate 22-8 as brown solid.
##STR00102##
[0237] Scheme 23 describes the synthesis of compound (I-SS). The
starting bromohydroxy compound ZZZZ1 was subjected to MOM
protection or SEM protection using MOMCl or SEMCl respectively in
the presence of a base to provide the product AAAAA1 which in turn
was formylated with DMF or N-formylpiperidine in the presence of
base like n-BuLi, s-BuLi, LDA, or LTMP at -78.degree. C. to give
product BBBBB1. The cross coupling reaction was done on BBBBB1 with
heteroaryl boronic acid or ester to provide the compound CCCCC1. In
one embodiment the heteroarylboron was 2-methylpyridine-4-boronic
acid. In another embodiment Pd.sub.2(dba).sub.3 in dioxane in the
presence of tripotassium phosphate and tricyclohexylphosphine was
used as a catalyst. The product CCCCC1 was deprotected in presence
of a Lewis acid to provide DDDDD1. In one embodiment the acid was
TFA. The compound DDDDD1 was treated with an amine
(R.sup.5--NH.sub.2) to provide imine EEEEE1. In one embodiment
imine formation was done in a mixed solvents of TFE and MeCN. The
imine EEEEE1 was allowed to react with a cyanide ion source to
provide the product (I-SS). In one embodiment the cyanide ion
source was TMSCN. In another embodiment the solvent was a mixture
of DCM-TFE.
##STR00103##
[0238] Scheme 23A described the synthesis of compound (I-TT).
2-Bromo-3-hydroxypyridine 23-1 was treated with MOMCl in the
presence of t-BuOK in THF resulting in to the formation of
2-bromo-3-(methoxymethoxy)pyridine 23-2. The MOM protected compound
underwent formylation with ethylformate in the presence of LDA at
in THF at -78.degree. C. to give
2-bromo-3-(methoxymethoxy)isonicotin aldehyde 23-3. The Suzuki
cross coupling reaction was done on 23-3 with
2-methylpyridine-4-boronic acid in the presence of tripotassium
phosphate, tricyclohexylphosphine and Pd.sub.2(dba).sub.3 in
dioxane to afford
3-(methoxymethoxy)-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde 23-4
which was treated with TFA-DCM solution to give MOM-de-protected
compound 23-5. The compound 23-5 was treated with amine
(R.sup.5NH.sub.2) to provide imine EEEEE2. In one embodiment imine
formation was done in a mixed solvent of TFE and MeCN. The imine
EEEEE2 was allowed to react with a cyanide ion source to provide
the product (I-TT). In one embodiment the cyanide ion source was
TMSCN. In another embodiment the solvent was a mixture of
DCM-TFE.
##STR00104##
[0239] Scheme 23B described the synthesis
N.sup.3-(3-chloro-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine 23-7. The 2-bromo-3-hydroxypyridine 23-1 was
treated with MOMCl in the presence of t-BuOK in THF resulting in to
the formation of 2-bromo-3-(methoxymethoxy)pyridine 23-2. The MOM
protected compound underwent formylation with ethylformate in the
presence of LDA in THF at -78.degree. C. to give
2-bromo-3-(methoxymethoxy)isonicotinaldehyde 23-3. The Suzuki cross
coupling reaction was done on 23-3 with 2-methylpyridine-4-boronic
acid in the presence of tripotassium phosphate,
tricyclohexylphosphine and Pd.sub.2(dba).sub.3 in dioxane to afford
3-(methoxymethoxy)-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde 23-4
which was treated with TFA-DCM solution to give MOM-de-protected
compound 23-5. The compound
3-hydroxy-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde 23-5 was
treated with 3-chloro-4-fluoroaniline in a mixed solvents of TFE
and MeCN to yield imine intermediate 23-6 which was further reacted
with TMSCN in a mixed solvents of DCM-TFE to afford desired product
N.sup.3-(3-chloro-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine 23-7 as a solid.
##STR00105##
[0240] Scheme 24 depicts the synthesis of (I-UU). The compound
BBBBB1 was coupled with a suitable substituted aryl- or heteroaryl
boronic acid or ester under cross-coupling reaction conditions to
provide compound FFFFF1. In one embodiment, the boronic ester used
was
N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
. In another embodiment, the coupling reaction was done in presence
of tripotassium phosphate, tricyclohexylphosphine and
Pd.sub.2(dba).sub.3 in dioxane. The compound FFFFF1 was deprotected
in presence of a Lewis acid to provide GGGGG1. In one embodiment,
the acid was TFA. The compound GGGGG1was next treated with an amine
(R.sup.5--NH.sub.2), a cyanide ion source and a Lewis acid in
sealed tube containing NH.sub.40Ac buffer solution to provide the
compound (I-UU). In one embodiment, the cyanide ion source was
TMSCN. In another embodiment, the Lewis acid was TMSOTf.
##STR00106##
[0241] Scheme 24A depicts the synthesis of compound (I-VV). The
compound 23-3 was coupled with
N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
under cross-coupling reaction conditions to provide 24-4. In one
embodiment, the coupling reaction was done in presence of
tripotassium phosphate, tricyclohexylphosphine and
Pd.sub.2(dba).sub.3 in dioxane. The compound 24-4 was deprotected
in presence of a Lewis acid to provide 24-5. In one embodiment the
acid was TFA. The compound 24-5 was treated with an amine
(R.sup.5--NH.sub.2), a cyanide ion source and a Lewis acid in
sealed tube containing NH.sub.4OAc buffer solution to provide the
compound (I-VV). In one embodiment the cyanide ion source was
TMSCN. In another embodiment the Lewis acid was TMSOTf.
##STR00107## ##STR00108##
[0242] Scheme 24B depicts the synthesis of 4- (2-amino-3-
((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-methylpicolina-
mide 24-6. The commercially available
4-chloro-pyridine-2-carboxylic acid 24-1 was treated with thionyl
chloride under refluxed to give intermediate 4-chloropicolinoyl
chloride which in-situ reacted with methyl amine in THF to afforded
amide 24-2. The compound 4-chloro-N-methylpicolinamide 24-2 was
reacted with bis(pinacolato)diboron under palladium catalyst to
give
N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
24-3. The compound 24-3 was coupled with
2-bromo-3-(methoxymethoxy)isonicotinaldehyde 23-3 (prepared
according to scheme 23B) in the presence of tripotassium phosphate,
tricyclohexylphosphine and Pd.sub.2(dba).sub.3 in dioxane to afford
4-formyl-3-(methoxymethoxy)-N-methyl[2,4'-bipyridine]-2'-carboxamide
24-4 which in turn underwent MOM-de-protection with TFA-DCM to form
4-formyl-3-hydroxy-N-methyl-[2,4'-bipyridine]-2'-carboxamide 24-5.
The compound 24-5 was coupled with 3-chloro-4-fluoroaniline to form
intermediate imine which was treated in-situ with TMSCN followed by
TMSOTf in a sealed tube containing NH.sub.4OAc buffer solution to
form cyclized product
4-(2-amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethylpicolinamide 24-6 as a solid.
##STR00109##
[0243] Scheme 25 describes the synthesis of compound (I-U). The
compound HHHHH1 was converted to a boronate IIIII1 by using a
borate reagent and a base. In one embodiment the borate used was
triisopropyl borate. In another embodiment the base used was
n-BuLi. The boronate IIIII1 was converted to hydroxyl compound
JJJJJ1 in presence of an oxidizing reagent. In one embodiment the
oxidizing reagent used was sodium perborate tetrahydrate in water.
The resulting compound JJJJJ1 was forwarded to MOM protection or
SEM protection using MOMCl or SEMCl respectively in the presence of
a base to provide the product VV1. The compound VV1 was formylated
with DMF or N-formylpiperidine in the presence of base like n-BuLi,
s-BuLi, LDA, or LTMP at -78.degree. C. to give product WW1. The
compound WW1 was deprotected in presence of a Lewis acid to provide
XX1. In one embodiment the acid was TFA. The compound XX1 was
treated with an amine (R.sup.5NH.sub.2) to provide the imine YY1.
The imine YY1 was treated with a cyanide ion source and a Lewis
acid to provide the compound (I-U). In one embodiment the cyanide
ion source was TMSCN. In another embodiment the Lewis acid was
TMSOTf.
##STR00110##
[0244] Scheme 25A describes the synthesis of compound (I-V). The
compound HHHHH.sub.2 was converted to a boronate IIIII2 by using a
borate reagent and a base. In one embodiment the borate used was
triisopropyl borate. In another embodiment the base used was
n-BuLi. The boronate IIIII2 was converted to hydroxyl compound
JJJJJ2 in presence of an oxidizing reagent. In one embodiment the
oxidizing reagent used was sodium perborate tetrahydrate in water.
The resulting compound JJJJJ2 was forwarded to MOM protection or
SEM protection using MOMCl or SEMCl respectively in the presence of
a base to provide the product VV2. The compound VV2 was formylated
with DMF or N-formylpiperidine in the presence of base like n-BuLi,
s-BuLi, LDA, or LTMP at -78.degree. C. to give product WW2. The
compound WW2 was deprotected in presence of a Lewis acid to provide
XX2. In one embodiment the acid was TFA. The compound XX2 was
treated with an amine (RNH.sub.2) to provide the imine YY2. The
imine YY2 was treated with a cyanide ion source and a Lewis acid to
provide the compound (I-V). In one embodiment the cyanide ion
source was TMSCN. In another embodiment the Lewis acid was
TMSOTf.
##STR00111##
[0245] Scheme 25B describe the synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine-2,3-dia-
mine 25-8. 2,6-Difluoropyridine 25-1 was treated with triisopropyl
borate in the presence of n-BuLi to give
2,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
25-2 which was further reacted with sodium perborate tetrahydrate
in water to yield 2.6-difluoro-3-hydroxypyridine 25-3. The compound
25-3 underwent MOM protection with MOMCl in the presence of
diisopropylamine to give MOM-protected compound 25-4. The
2,6-difluoro-3-(methoxymethoxy)pyridine 25-4 was formylated with
DMF in the presence of n-BuLi to afford
2,6-difluoro-3-(methoxymethoxy)isonicotinaldehyde 25-5 which was
deprotected by TFA-DCM to give 25-6. The
2,6-difluoro-3-hydroxyisonicotinaldehyde 25-6 was coupled with
3-chloro-4-fluoroaniline to give imine 25-7 which was further
treated with TMSCN and TMSOTf resulting into the formation of first
Strecker product which in-situ underwent intramolecular cyclization
to afford
N.sup.3-(3-chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine-2,3-dia-
mine 25-8 as brown solid.
##STR00112##
[0246] Scheme 26 depicts the synthesis of compound (I-YY). The
compound AAAAA1 was converted to OOOOO1 by using an alkyl or
aromatic amine under cross-coupling reaction conditions. In one
embodiment Buchwald-Hartwig amination reaction was performed as
cross-coupling reaction. In another embodiment diphenylamine was
used an aryl amine. In yet another embodiment the catalysts used
were Pd.sub.2(dba).sub.3 and Dppf. The compound OOOOO1 underwent
formylation with DMF or N-formylpiperidine in the presence of base
like n-BuLi, s-BuLi, LDA, or LTMP at -78.degree. C. to give product
PPPPP1. The compound PPPPP1 was deprotected in presence of a Lewis
acid to provide QQQQQ1. In one embodiment the acid was TFA. The
compound QQQQQ1 was treated with an amine (R.sup.5--NH.sub.2) to
provide imine RRRRR1. The amine RRRRR1 was treated with a cyanide
ion source and a Lewis acid to provide the compound (I-YY). In one
embodiment the cyanide ion source was TMSCN. In another embodiment
the Lewis acid was TMSOTf.
##STR00113##
[0247] Scheme 26A depicts the synthesis of compound (I-ZZ). The
Buchwald-Hartwig amination reaction was done on
2-bromo-3-(methoxymethoxy)pyridine 23-2 with diphenyl amine in the
presence of Pd.sub.2(dba).sub.3 and Dppf to afford
3-(methoxymethoxy)-N,N-diphenylpyridin-2-amine 26-1. The compound
26-1 was formylated with DMF in the presence of n-BuLi to give
2-(diphenylamino)-3-(methoxymethoxy)isonicotinaldehyde 26-2 which
underwent MOM-deprotection to yielded
2-(diphenylamino)-3-hydroxyisonicotinaldehyde 26-3. The compound
26-3 was treated with amine (R.sup.5--NH.sub.2) to provide imine
RRRRR2. The amine RRRRR2 was treated with a cyanide ion source and
a Lewis acid to provide the compound (I-ZZ). In one embodiment the
cyanide ion source was TMSCN. In another embodiment the Lewis acid
was TMSOTf.
##STR00114##
[0248] Scheme 26B depicts the synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-N.sup.7,N.sup.7-diphenylfuro[2,3-c]pyri-
dine-2,3,7-triamine 26-5. The Buchwald-Hartwig amination reaction
was performed on 2-bromo-3-(methoxymethoxy)pyridine 23-2 with
diphenylamine in the presence of Pd.sub.2(dba).sub.3 and Dppf to
afford 3-(methoxymethoxy)-N,N-diphenylpyridin-2-amine 26-1. The
compound 26-1 was formylated with DMF in the presence of n-BuLi to
give 2-(diphenylamino)-3-(methoxymethoxy)isonicotinaldehyde 26-2.
Compound 26-2h underwent MOM-deprotection to yield
2-(diphenylamino)-3-hydroxyisonicotinaldehyde 26-3. The compound
26-3 was coupled with 3-chloro-4-fluoroaniline to form an imine
26-4. The immune 26-4 was further treated with TMSCN in TFE
resulting into the formation of a first Strecker product which
underwent in-situ intramolecular cyclization to form
N.sup.3-(3-chloro-4-fluorophenyl)-N.sup.7,N.sup.7-diphenylfuro[2,3-c]pyri-
dine-2,3,7-triamine 26-5 as a solid.
##STR00115##
[0249] Scheme 27 depicts the preparation of compound (I-CCC). The
starting compound SSSSS1 was treated with a base and then allowed
to react with a suitable alkyl halide to form a compound TTTTT1.
The compound TTTTT1 was converted to a compound UUUUU1 under acid
catalysis. In one embodiment, PPA in chlorobenzene was used as an
acid catalyst. The compound UUUUU1 was halogenated to provide a
compound VVVVV1. In one embodiment, NBS, in a mixture chloroform
and acetic acid, was used for halogenation. The compound VVVVV1 was
nitrated to provide a nitro compound WWWWW1. In one embodiment,
nitric acid was used for nitration. The compound WWWWW1 was treated
with an amine (R.sup.5--NH.sub.2) to provide a compound XXXXX1. The
compound XXXXX1 was reacted with an aryl or heteroaryl boronic acid
or ester in a cross-coupling reaction to provide a compound YYYYY1.
In one embodiment, a mixture of DMF and water was used a solvent.
In another embodiment, Pd(PPh.sub.3).sub.4 and an inorganic base
were used as catalysts. In another embodiment, the inorganic base
was any of K.sub.2CO.sub.3, KHCO.sub.3, or CsF or K.sub.3PO.sub.4.
In yet another embodiment, the boronic acid was
2-methyl-4-pyridinylboronic acid. Next, nitro compound YYYYY1 was
reduced to an amine to provide a compound (I-CCC). In one
embodiment, Pd/C and hydrogen were used for reduction.
##STR00116##
[0250] Scheme 27A describes the preparation of compound (I-DDD).
Compound 2-bromothiophenol was treated with potassium carbonate in
acetone and then allowed to react with bromoacetaldehyde
diethylacetal to form (2-bromophenyl)(2,2-diethoxyethyl)sulfane
27-2. The compound 27-2 was allowed to react with PPA in
chlorobenzene to provide 7-bromobenzo[b]thiophene 27-3. Compound
27-3 in turn was brominated to form 3,7-dibromobenzo[b]thiophene
27-4 by using NBS in a mixture chloroform and acetic acid. This was
followed by nitration of 27-4 to provide
3,7-dibromo-2-nitrobcnzo[b]thiophene (27-5). The compound 27-5 was
treated with an amine (R.sup.5--NH.sub.2) to provide a compound
XXXXX2. The compound XXXXX2 was reacted with an optionally
substituted aryl or heteroaryl boronic acid or ester in a
cross-coupling reaction to provide a compound YYYYY2. In one
embodiment, a mixture of DMF and water was used a solvent. In
another embodiment, Pd(PPh.sub.3).sub.4 and an inorganic base were
used as catalysts. In another embodiment, the inorganic base was
any one of K.sub.2CO.sub.3, KHCO.sub.3, CsF or K.sub.3PO.sub.4. In
yet another embodiment, the boronic acid was
2-methyl-4-pyridinylboronic acid. Next, nitro compound YYYYY2 was
reduced to a amine to provide a compound (I-DDD). In one
embodiment, Pd/C and hydrogen were used for reduction.
##STR00117##
[0251] Scheme 27B describes the preparation of compound
N.sup.3-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)benzo[b]thiophene-2,3-di-
amine 27-8. 2-bromothiophenol was treated with potassium carbonate
in acetone and then allowed to react with bromoacetaldehyde
diethylacetal to form (2-bromophenyl)(2,2-diethoxyethyl)sulfane
27-2. The compound 27-2 was allowed to react with PPA in
chlorobenzene to form 7-bromobenzo[b]thiophene 27-3. The compound
27-3 in turn was brominated to form 3,7-dibromobenzo[b]thiophene
27-4 by using NBS in a mixture chloroform and acetic acid. This was
followed by nitration of compound 27-4 to provide
3,7-dibromo-2-nitrobenzo[b]thiophene 27-5. The compound 27-5 was
treated with 3-chloroaniline in DMF to provide
7-bromo-N-(3-chlorophenyl)-2-nitrobenzo[b]thiophen-3-amine 27-6.
The compound 27-6 was reacted with 2-methyl-4-pyridinylboronic acid
in a mixture of DMF and water in the presence of K.sub.3PO.sub.4
and Pd(PPh.sub.3).sub.4 to form
N-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)-2-nitrobenzo[b]thiophen-3-ami-
ne 27-7. Next, nitro compound 27-7 was reduced to an amine by using
Pd/C and hydrogen to form
N.sup.3-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)benzo[b]thiophene-2,3-di-
amine 27-8.
##STR00118##
[0252] Scheme 28 depicts the synthesis of compound (1-U). Compound
EEEE1-1 was coupled with arylboronic acid under Suzuki
cross-coupling reaction condition to form compound SS1. In one
embodiment, a mixture of DMF and water was used as a solvent. In
another embodiment, the solvent was 1,4-dioxane. In yet another
embodiment, Pd(PPh.sub.3).sub.4 and an inorganic base were used as
catalysts. In still another embodiment, the catalyst was
Pd.sub.2(dba).sub.3. In a further embodiment, the inorganic base
was any of K.sub.2CO.sub.3, KHCO.sub.3, CsF or K.sub.3PO.sub.4. In
yet another embodiment, the arylboronic acid was phenylboronic
acid. The compound SS1 was treated with iodine in basic medium to
provide compound TT1. In one embodiment, the base was
Na.sub.2CO.sub.3. The compound TT1 was MOM-protected with MOMCl in
the presence of a base to form a MOM-protected compound UU1. In one
embodiment, the base was potassium tert-butoxide. The compound UU1
was added to a freshly prepared sodium alkoxide or aryloxide
solution. Next, to the resulting mixture was addedCuBr to form a
compound VV1. In one embodiment, the sodium aryloxide was sodium
phenoxide. The compound VV1 was formylated with DMF in the presence
of any on or more or n-BuLi, s-BuLi, LDA, and TMEDA to form
compound WW1. Compound WW1 next under went deprotection under
acidic conditions to yield a compound XX1. In one embodiment, the
acid was HCl. In another embodiment the acid was TFA. Compound XX1
was first coupled with an amine (R.sup.5--NH.sub.2) and then
treated with trialkylsilyl cyanide. The resulting mixture was next
treated with a Lewis acid to form compound (I-U) as a solid. In one
embodiment, the cyanide source was TMSCN. In another embodiment the
cyanide source was NaCN. In another embodiment, the Lewis acid was
any of TMSOTf, Sc(OTO.sub.3, Fe(OTf).sub.2, Ni(OTf).sub.2 or
In(OTf).sub.3.
##STR00119##
[0253] Scheme 28A describes the synthesis of compound (I-V).
2-Iodo-3-hydroxypyridine 14-2 was coupled with arylboronic acid
under Suzuki cross coupling reaction condition to form compound B2.
In one embodiment, a mixture of DMF and water was used a solvent.
In another embodiment, Pd(PPh.sub.3).sub.4 and an inorganic base
were used as catalysts. In still another embodiment, the inorganic
base was any of K.sub.2CO.sub.3, KHCO.sub.3, CsF, or
K.sub.3PO.sub.4. In yet another embodiment, the arylboronic acid
was phenylboronic acid. Compound B2 was treated with iodine in
basic medium to provide compound TT2. In one embodiment, the base
used in the basic medium was Na.sub.2CO.sub.3. The compound TT2 was
MOM-protected with MOMCl in the presence of potassium tert-butoxide
to form a MOM-protected compound UU2. The compound UU2 was added to
freshly prepared sodium aryloxide solution. Next, to the reaction
mixture was added CuBr to form a compound VV2. In one embodiment,
the sodium aryloxide was sodium phenoxide. The compound VV2 was
formylated with DMF in the presence of any of n-BuLi, s-BuLi, or
LDA, and TMEDA to form compound WW2. Next, compound WW2 underwent
deprotection under acidic conditions to yield a compound XX2. In
one embodiment, the acid used was HCl. The compound XX2 was coupled
with an amine (R.sup.5--NH.sub.2) and then treated first with
trialkylsilyl cyanide and then with a Lewis acid to form a compound
(I-V) as a solid. In one embodiment, the cyanide source was TMSCN.
In another embodiment, the lewis acid was TMSOTf.
##STR00120##
[0254] Scheme 28B describes the synthesis of compound
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenoxy-7-phenylfuro[2,3-c]pyridine-2-
,3-diamine 28-4.2-Iodo-3-hydroxypyridine 14-2 was coupled with
phenylboronic acid under Suzuki cross-coupling reaction to form
2-Phenyl-3-hydroxypyridine 14-3. Compound 14-3 was treated with
iodine in basic medium to provide 6-iodo-2-phenylpyridin-3-ol 14-4
which underwent MOM-protection with MOMCl in the presence of
potassium tert-butoxide to form a MOM-protected compound
6-iodo-3-(methoxymethoxy)-2-phenylpyridine 14-5. The compound 14-5
was added to freshly prepared sodium phenoxide solution followed by
the addition of CuBr. The resulting reaction mixture was refluxed
for 16 h forming 3-(methoxymethoxy)-6-phenoxy-2-phenylpyridine
28-1. The compound 28-1 was formylated with DMF in the presence of
n-BuLi and TMEDA to form
3-(methoxymethoxy)-6-phenoxy-2-phenylisonicotinaldehyde 28-2.
Compound 28-2 underwent deprotection to yield
3-hydroxy-6-phenoxy-2-phenylisonicotinaldehyde 28-3. The compound
28-3 was first coupled with 4-fluoro-3-chloroaniline and then
treated with TMSCN. The resulting mixture was next treated with
TMSOTf resulting in the formation of
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenoxy-7-phenylfuro[2,3-c]pyridine-2-
,3-diamine 28-4 as a solid.
[0255] Accordingly, the invention also relates to a method of
preparing a compound of formula (I-C), said method comprising:
##STR00121##
[0256] wherein: [0257] X.sup.1 is CR.sup.1, N, or NO; [0258]
X.sup.2 is CR.sup.2, N, or NO; [0259] X.sup.3 is CR.sup.3, N, or
NO; [0260] X.sup.4 is CR.sup.4, N, or NO; and [0261] R.sup.5 is
selected from the group consisting of H, optionally substituted
C.sup.1-C.sup.6 alkyl, optionally substituted mono or bicyclic
C.sup.6-C.sup.14 aryl, optionally substituted mono or bicyclic
heteroaryl, optionally substituted (aryl)alkyl, optionally
substituted mono or bicyclic cycloalkyl, optionally substituted
mono or bicyclic heterocyclyl, C.sub.1-C.sub.6haloalkyl, optionally
substituted hcterocyclyl(alkyl), optionally substituted
heteroaryl(alkyl), hydroxyalkyl, and perfluoroalkyl;
[0262] (i) reacting
##STR00122##
with an amine (R.sup.5--NH.sub.2); [0263] (ii) reacting the product
of step (i) with a cyanide salt; and [0264] (iii) reacting the
product of step (ii) with a Lewis acid.
[0265] In a further embodiment, the Lewis acid is trimethylsilyl
trifluoromethanesulfonate, Sc(OTf).sup.3, Fe(OTf).sup.2,
Ni(OTf).sup.2, or In(OTf).sup.3. In a still further embodiment, the
cyanide salt is selected from group consisting of a trialkyl silyl
cyanide, NaCN, KCN, and Zn(CN).sup.2. In yet another embodiment,
the trialkyl silyl cyanide is TMSCN. In a still further embodiment,
the reaction of step (iii) is performed in the presence of a buffer
solution. In one embodiment, the buffer solution is ammonium
acetate buffer.
[0266] Also falling within the scope of the invention are compounds
that are obtainable by the practice of methods disclosed herein. In
an embodiment, the invention relates to a compound obtainable by a
method of preparing a compound of formula (I-C), said method
comprising:
##STR00123##
[0267] wherein: [0268] X.sup.1 is CR.sup.1, N, or NO; [0269]
X.sup.2 is CR.sup.2, N, or NO; [0270] X.sup.3 is CR.sup.3, N, or
NO; [0271] X.sup.4 is CR.sup.4, N, or NO; and [0272] R.sup.5 is
selected from the group consisting of H, optionally substituted
C.sup.1-C.sup.6 alkyl, optionally substituted mono or bicyclic
C.sup.6-C.sup.14 aryl, optionally substituted mono or bicyclic
heteroaryl, optionally substituted (aryl)alkyl, optionally
substituted mono or bicyclic cycloalkyl, optionally substituted
mono or bicyclic heterocyclyl, C.sup.1-C.sup.6 haloalkyl,
optionally substituted heterocyclyl(alkyl), optionally substituted
heteroaryl(alkyl), hydroxyalkyl, and perfluoroalkyl;
[0273] (i) reacting
##STR00124##
with an amine (R.sup.5--NH.sub.2); [0274] (ii) reacting the product
of step (i) with a cyanide salt; and [0275] (iii) reacting the
product of step (ii) with a Lewis acid.
[0276] In a further embodiment, the Lewis acid is trimethylsilyl
trifluoromethanesulfonate, Sc(OTf).sup.3, Fe(OTf).sup.2,
Ni(OTf).sup.2, or In(OTf).sup.3. In a still further embodiment, the
cyanide salt is selected from group consisting of a trialkyl silyl
cyanide, NaCN, KCN, and Zn(CN).sup.2. In yet another embodiment,
the trialkyl silyl cyanide is TMSCN. In a still further embodiment,
the reaction of step (iii) is performed in the presence of a buffer
solution. In one embodiment, the buffer solution is ammonium
acetate buffer.
[0277] In still another embodiment, invention relates to
intermediate compounds that are products of one or more of steps
(i) (iii) of the method of preparing a compound of formula (I-C).
In one embodiment, the product or intermediate of step (i) is an
imine.
Pharmaceutical Compositions
[0278] Pharmaceutical compositions useful herein contain a compound
of formula (I) or metabolites thereof, or prodrugs thereof
("compounds of the invention" or "compounds") in a pharmaceutically
acceptable carrier optionally with other pharmaceutically inert or
inactive ingredients. In another embodiment, a compound of formulae
(I)-(IV) is present in a single composition. In another embodiment,
a metabolite of a compound of formulae (I)-(IV) is present in a
single composition. In yet another embodiment, a prodrug of a
compound of formula (I), including without limitation, a compound
having any one of the formulae (II)-(IV) is present in a single
composition. In a further embodiment, a compound of formula (I) or
a metabolite thereof, or a pharmaceutically accetable salt or
prodrug thereof is combined with one or more excipients and/or one
or more of other therapeutic agents as described below.
[0279] Pharmaceutical compositions comprise an amount of a compound
of formula (I) or a metabolite thereof, or prodrug thereof, or
pharmaceutically acceptable salt thereof that is effective for
regulating one or more of indoleamine 2,3-dioxygenase-1 or
indoleamine 2,3-dioxygense-2 or tryptophan 2,3-dioxygenase enzymes.
Pharmaceutical compositions comprise an amount of compound of
formula (I) or a metabolite thereof, or a pharmaceutical salt or
prodrug thereof that is effective for regulating the kynurenine
pathway. The pharmaceutical compositions useful herein comprise an
amount of a compound of formula (I) or a metabolite thereof, or
prodrug thereof, or a pharmaceutically acceptable salt thereof that
is effective for regulating the kynurenine pathway by inhibiting
one or more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes in a
subject. The pharmaceutical compositions comprise an amount of a
compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof that is
effective for regulating one or more of indoleamine
2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase enzyme in a subject. In one aspect, the
pharmaceutical compositions comprise an amount of a compound or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof that is effective for reducing kynurenine pathway
metabolites and/or altering (for example increasing) tryptophan
levels and/or reducing kynurenine/tryptophan ratio in a subject.
The pharmaceutical compositions comprise an amount of a compound of
Formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof that is effective for reducing
or eliminating autoimmune antibody in a subject. In another aspect,
pharmaceutical compositions comprise an amount of a compound of
formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof that is effective for reducing
immune suppression in a subject.
[0280] Specifically, the dosage of the compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof to achieve a therapeutic effect will depend on the
formulation, age, weight and sex of the patient and route and
frequency of delivery. It is also contemplated that the treatment
and dosage of the compound of formula (I) or a metabolite thereof,
or a pharmaceutically acceptable salt or prodrug thereof may be
administered in unit dosage form and that one skilled in the art
would adjust the unit dosage form accordingly to reflect the
relative level of activity. The decision as to the particular
dosage to be employed (and the number of times to be administered
per day) is within the discretion of the ordinarily-skilled
physician, and may be varied by titration of the dosage to the
particular circumstances to produce the desired therapeutic effect.
In one embodiment, the therapeutically effective amount is about
0.01 mg/kg to 10 mg/kg body weight. In another embodiment, the
therapeutically effective amount is less than about 5 g/kg, about
500 mg/kg, about 400 mg/kg, about 300 mg/kg, about 200 mg/kg, about
100 mg/kg, about 50 mg/kg, about 25 mg/kg, about 10 mg/kg, about 1
mg/kg, about 0.5 mg/kg, about 0.25 mg/kg, about 0.1 mg/kg, about
100 .mu.g/kg, about 75 .mu.g/kg, about 50 .mu.g/kg, about 25
.mu.g/kg, about 10 .mu.g/kg, or about 1 .mu.g/kg. However, the
therapeutically effective amount of the compound of formula (I) or
a metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof can be determined by the attending physician and
depends on the condition or disease treated, the compound
administered, the route and frequency of delivery, the age, weight,
severity of the patient's symptoms and response pattern of the
patient.
[0281] The therapeutically effective amounts may be provided on
regular schedule, i.e., daily, weekly, monthly, or yearly basis or
on an irregular schedule with varying administration days, weeks,
months, etc. Alternatively, the therapeutically effective amount to
be administered may vary. In one embodiment, the therapeutically
effective amount for the first dose is higher than the
therapeutically effective amount for one or more of the subsequent
doses. In another embodiment, the therapeutically effective amount
for the first dose is lower than the therapeutically effective
amount for one or more of the subsequent doses. Equivalent dosages
may be administered over various time periods including, but not
limited to, about every 2 hours, about every 6 hours, about every 8
hours, about every 12 hours, about every 24 hours, about every 36
hours, about every 48 hours, about every 72 hours, about every
week, about every two weeks, about every three weeks, about every
month, about every two months, about every four months, about every
six months, about every 9 months, and about every year. The number
and frequency of dosages corresponding to a completed course of
therapy will be determined according to the judgment of a
health-care practitioner. The therapeutically effective amounts
described herein refer to total amounts administered for a given
time period; that is, if more than one compound of formula (I) or a
metabolite thereof, or prodrug thereof, or a pharmaceutically
acceptable salt thereof is administered, the therapeutically
effective amounts correspond to the total amount administered.
[0282] The pharmaceutical compositions containing a compound of
formula (I) may be formulated neat or with one or more
pharmaceutical carriers for administration. The amount of the
pharmaceutical carrier(s) is determined by the solubility and
chemical nature of the compound of formula (I) or a metabolite
thereof, or prodrug thereof, chosen route of administration and
standard pharmacological practice. The pharmaceutical carrier(s)
may be solid or liquid and may incorporate both solid and liquid
carriers. A variety of suitable liquid carriers are known and may
be readily selected by one of skill in the art. Such carriers may
include, for example, DMSO, saline, buffered saline,
hydroxypropylcyclodextrin, and mixtures thereof. Similarly, a
variety of solid carriers and excipients are known to those of
skill in the art. The compounds of formula (I) may be administered
by any route, taking into consideration the specific condition or
disease for which it has been selected. The compounds of formula
(I) or a metabolite thereof, or prodrug thereof may, be delivered
orally, by injection, inhalation (including orally, intranasally
and intratracheally), ocularly, transdermally, intravascularly,
subcutaneously, intramuscularly, sublingually, intracranially,
epidurally, rectally, and vaginally, among others.
[0283] Although the compound of formula (I) or a metabolite
thereof, or a pharmaceutically salt thereof or prodrug thereof may
be administered alone, it may also be administered in the presence
of one or more pharmaceutical carriers that arc physiologically
compatible. The carriers may be in dry or liquid form and must be
pharmaceutically acceptable. Liquid pharmaceutical compositions are
typically sterile solutions or suspensions. When liquid carriers
are utilized for parenteral administration, they are desirably
sterile liquids. Liquid carriers are typically utilized in
preparing solutions, suspensions, emulsions, syrups and elixirs. In
one embodiment, the compound of formula (I) or a metabolite
thereof, or a pharmaceutically salt thereof or prodrug thereof is
dissolved a liquid carrier. In another embodiment, the compound of
formula (I) or a metabolite thereof, or a pharmaceutically salt
thereof or prodrug thereof is suspended in a liquid carrier. One of
skill in the art of formulations would be able to select a suitable
liquid carrier, depending on the route of administration. The
compound of formula (I) or a metabolite thereof, or a
pharmaceutically salt thereof or prodrug thereof may alternatively
be formulated in a solid carrier. In one embodiment, the
composition may be compacted into a unit dose form, i.e., tablet or
caplet. In another embodiment, the composition may be added to unit
dose form, i.e., a capsule. In a further embodiment, the
composition may be formulated for administration as a powder. The
solid carrier may perform a variety of functions, i.e., may perform
the functions of two or more of the excipients described below. For
example, solid carrier may also act as a flavoring agent,
lubricant, solubilizer, suspending agent, filler, glidant,
compression aid, binder, disintegrant, or encapsulating
material.
[0284] The composition may also be sub-divided to contain
appropriate quantities of the compound of formula (I) or a
metabolite thereof, or a pharmaceutically salt thereof or prodrug
thereof. For example, the unit dosage can be packaged compositions,
e.g., packeted powders, vials, ampoules, prefilled syringes or
sachets containing liquids.
[0285] Examples of excipients which may be combined with one or
more compound of formula (I) or a metabolite thereof, or prodrug
thereof include, without limitation, adjuvants, antioxidants,
binders, buffers, coatings, coloring agents, compression aids,
diluents, disintegrants, emulsifiers, emollients, encapsulating
materials, fillers, flavoring agents, glidants, granulating agents,
lubricants, metal chelators, osmo-regulators, pH adjustors,
preservatives, solubilizers, sorbents, stabilizers, sweeteners,
surfactants, suspending agents, syrups, thickening agents, or
viscosity regulators. See, for example, the excipients described in
the "Handbook of Pharmaceutical Excipients", 5.sup.th Edition,
Eds.: Rowe, Sheskey, and Owen, APhA Publications (Washington,
D.C.), Dec. 14, 2005, which is incorporated herein by
reference.
[0286] In one embodiment, the compositions may be utilized as
inhalants. For this route of administration, compositions may be
prepared as fluid unit doses using a compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof, and a vehicle for delivery by an atomizing spray
pump or by dry powder for insufflation.
[0287] In another embodiment, the compositions may be utilized as
aerosols, i.e., oral or intranasal. For this route of
administration, the compositions are formulated for use in a
pressurized aerosol container together with a gaseous or liquefied
propellant, e.g., dichlorodifluoromethane, carbon dioxide,
nitrogen, propane, and the like. Also provided is the delivery of a
metered dose in one or more actuations.
[0288] In another embodiment, the compositions may be administered
by a sustained delivery device. "Sustained delivery" as used herein
refers to delivery of a compound of formula (I) or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
which is delayed or otherwise controlled. Those of skill in the art
know suitable sustained delivery devices. For use in such sustained
delivery devices, the compound of formula (I) or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
is formulated as described herein.
Labeled Compounds and Assay Methods
[0289] Another aspect of the present invention relates to
fluorescent dyes, spin label, heavy metal, or isotopically- or
radio-labeled compounds of the invention that would be useful not
only in imaging but also in assays both in vitro and in vivo, for
localizing and quantitating one or more of IDO1 or IDO2 or TDO
enzymes in blood or tissue samples of mammals or in cells, and for
identifying or screening for ligands of one or more of IDO1 or IDO2
or TDO by inhibition binding of a labeled compound. Compounds of
the invention may also be conjugated to other therapeutics or assay
reagents, for example, to biotherapeutics such as targeted
antibodies or antibody fragments, or drug targets or antibodies,
antibody fragment or a protein as reagents for research or test
purposes or for any other purpose. Accordingly, the present
invention includes enzyme assays for one or more of IDO1 or IDO2 or
TDO enzymes that contain such labeled compounds. Examples of assays
include without limitation ELISA, RTA, ELTSPOT etc.
[0290] One aspect of the invention includes isotopically-labeled
compounds, which are identical to those shown in formula (I) or
metabolites thereof, or pharmaceutically acceptable salts or
prodrugs thereof, but for the fact that one or more atoms are
replaced by an atom having an atomic mass or mass number usually
different from the atomic mass or mass number usually found in
nature. Examples of isotopes that can be incorporated into
compounds of the invention include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such
as .sup.2H, .sup.3H, .sup.13C, .sup.15N, .sup.18O, .sup.17O,
.sup.31P, .sup.32P, .sup.35S, .sup.18F, and .sup.36Cl,
respectively. Compounds of the invention which contain the
aforementioned isotopes and other isotopes of other atoms are
within the scope this of invention.
[0291] Synthetic methods for incorporating radio-isotopes into
organic compounds are applicable to compounds of the invention and
are well-known in the art. The radionucliide that is incorporated
in the instant radio-labeled compounds will depend on the specific
application of that radiolabeled compound. It will be understood
that, in a compound where such isotopic substitution is made, the
following atoms, where present, may vary, so that for example, any
hydrogen may be .sup.2H/D, any carbon may be .sup.13C, or any
nitrogen may be .sup.15N, and that the presence and placement of
such atoms may be determined by a person having skill in the art.
For example, for an IDO1, IDO2, or TDO enzyme labeling and
competition assays, compounds that are incorporate .sup.2H/D,
.sup.3H, .sup.14C .sup.82Br, .sup.125I, .sup.131I, .sup.35S will
generally be most useful. The radioactive isotopes .sup.3H, and
.sup.14C, are particularly useful for this purpose in view of their
ease of incorporation and ready means of detection.
[0292] Likewise, the compounds of the invention may include the
preparation of isotopic variants with radioisotopes, in the
instance for example, where the resulting compounds may be used for
drug and/or substrate tissue distribution studies. For example, in
radio-imaging applications .sup.11C, .sup.18F, .sup.125I,
.sup.123I, .sup.124I, .sup.131I, .sup.75Br, .sup.77Br will
generally be most useful. Further, compounds may be prepared that
arc substituted with positron emitting isotopes, such as .sup.11C,
.sup.18F, .sup.15O, and .sup.13N, and would be useful in Positron
Emission Topography (PET) studies for examining substrate receptor
occupancy. In some embodiments, the radionuclide is selected from a
group of .sup.3H, .sup.14C, .sup.125I, .sup.35S, .sup.82Br.
Tritiated .sup.3H and carbon 14 i.e., .sup.14C, are preferred for
their ease of preparation and detectability. Further, substitution
with heavier isotopes such as deuterium, i.e. .sup.2H/D, can afford
certain therapeutic advantages resulting from greater metabolic
stability, for example, increased in vivo half life or reduced
dosage requirements, and hence can be preferred in some
circumstances. All isotopic variants of the compounds provided
herein, radioactive or not, are intended to be encompassed within
the scope of the invention.
[0293] A isotopically- or radio-labeled compound can be used in a
screening assay to identify or evaluate compounds or drug targets.
In general terms, a newly synthesized or identified compound (i.e.,
test compound) can be evaluated for its ability to reduce binding
of the radio-labeled compound to one or more of IDO1 or IDO2 or TDO
enzymes. Accordingly, the ability of a test compound to compete
with an labeled compound such as isotopically including
radio-labeled compound or fluroscein-labeled compound for binding
to one or more of IDO1, IDO2, or TDO enzymes directly correlates to
its binding activity.
Articles Of Manufacture
[0294] Also provided herein are kits or packages of pharmaceutical
formulations containing the compounds of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof, or compositions described herein. The kits may be
organized to indicate a single formulation or combination of
formulations to be taken at each desired time.
[0295] Suitably, the kit contains packaging or a container with the
compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof formulated for
the desired delivery route. Suitably, the kit contains instructions
on dosing and an insert regarding the active agent. Optionally, the
kit may further contain instructions for monitoring circulating
levels of product and materials for performing such assays
including, e.g., reagents, well plates, containers, markers or
labels, and the like. Such kits are readily packaged in a manner
suitable for treatment of a desired indication. For example, the
kit may also contain instructions for use of a spray pump or other
delivery device. Other suitable components to include in such kits
will be readily apparent to one of skill in the art, taking into
consideration the desired indication and the delivery route.
[0296] The compounds of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof, or
compositions described herein can be a single dose or for
continuous or periodic discontinuous administration. For continuous
administration, a package or kit can include the compound of
formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof, in each dosage unit (e.g.,
solution, lotion, tablet, pill, or other unit described above or
utilized in drug delivery), and optionally instructions for
administering the doses daily, weekly, or monthly, for a
predetermined length of time or as prescribed. When the compound of
formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof is to be delivered periodically
in a discontinuous fashion, a package or kit can include placebos
during periods when the compound of formula (I) or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
is not delivered. When varying concentrations of a composition, of
the components of the composition, or the relative ratios of the
compounds of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof, or therapeutic
agents within a composition over time is desired, a package or kit
may contain a sequence of dosage units which provide the desired
variability.
[0297] A number of packages or kits are known in the art for
dispensing pharmaceutical agents for periodic oral use. In one
embodiment, the package has indicators for each period. In another
embodiment, the package is a labeled blister package, dial
dispenser package, or bottle.
[0298] The packaging means of a kit may itself be geared for
administration, such as an inhalant, syringe, pipette, eye dropper,
or other such apparatus, from which the formulation may be applied
to an affected area of the body, such as the lungs, injected into a
subject, or even applied to and mixed with the other components of
the kit.
[0299] The compositions of these kits also may be provided in dried
or lyophilized forms. When reagents or components are provided as a
dried form, reconstitution generally is by the addition of a
suitable solvent. It is envisioned that the solvent also may be
provided in another package.
[0300] The kits of the present invention also will typically
include a means for containing the vials in close confinement for
commercial sale such as, e.g., injection or blow-molded plastic
containers into which the desired vials are retained. Irrespective
of the number or type of packages and as discussed above, the kits
also may include, or be packaged with a separate instrument for
assisting with the injection/administration or placement of the
composition within the body of an animal. Such an instrument may be
an inhalant, syringe, pipette, forcep, measuring spoon, eye dropper
or any such medically approved delivery means.
[0301] In one embodiment, a kit is provided and contains a compound
of formula (I). In another embodiment the kit comprises a compound
of formula (I) or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof. In yet another embodiment, the
kit comprises a metabolite of a compound of formulae (I)-(IV). In
yet another embodiment, the kit contains a prodrug of a compound of
formulae (I)-(IV). The compound of formula (I) or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
may be in the presence or absence of one or more of the carriers or
excipients described above. In still another embodiment, the kit
contains a compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof may be in one
or more of other therapeutic agents as described herein. The kit
may optionally contain instructions for administering the
medication and the compound of formula (I) or a metabolite thereof,
or a pharmaceutically acceptable salt or prodrug thereof to a
subject having a disease characterized by (i) the dysregulation of
the kynurenine pathway caused by dysregulated indoleamine
2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2 and/or
tryptophan 2,3-dioxygenase activity, or (ii) immune suppression or
(iii) autoimmunity, or (iv) increased kynurenine metabolites or (v)
decreased tryptophan or (vi) increased kynurenine/tryptophan ratio
or (vii) with inflammation.
[0302] In a further embodiment, a kit is provided and contains a
compound of formula (I) or a metabolite thereof, or prodrug
thereoof in a second dosage unit, and one or more of the carriers
or excipients described above in a third dosage unit. The kit may
optionally contain instructions for administering the medication
and the compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to a subject
having a disease characterized by (i) abnormal immune suppression
resulting from dysregulation of the kynurenine pathway, or (ii)
autoimmunity, or (iii) increased kynurenine metabolites, or (iv)
decreased tryptophan, or (v) increased kynurenine/tryptophan
ratio.
[0303] In a further embodiment, a kit is provided and contains a
compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof in a second
dosage unit, and one or more of the carriers or excipients
described above in a third dosage unit. The kit may optionally
contain instructions for administering the medication and the
compound of formula (I) or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to a subject
having a disease characterized by abnormal immune suppression
resulting from enzymatic activity of indoleamine 2,3-dioxygenase-1
and/or indoleamine 2,3-dioxygenase-2 and/or tryptophan
2,3-dioxygenase.
Methods of Use
[0304] The compounds of formula (I) or metabolites thereof, or a
pharmaceutically acceptable salt or prodrugs thereof, and
pharmaceutical compositions described herein are useful in treating
or regulating diseases or conditions associated with kynurenine
pathway. Specifically, the compounds are useful in treating or
regulating diseases or conditions associated with increased
kynurenine pathway metabolites, for e.g., kynurenine or altered
(for example, decreased) tryptophan levels. The compounds are
useful for the treatment of disease or condition associated with
one or more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-disoxygenase-2 or tryptophan 2,3-dioxygenase enzymes. The
compounds of the invention are useful in the treatement of immune
suppression. In one aspect, the immune suppression is associated
with one or more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-disoxygenase-2 or tryptophan 2,3-dioxygenase enzymes. The
compounds and pharmaceutical compositions described herein are
useful in regulating diseases which are associated with increased
immune suppression resulting from dysregulation of the kynurenine
pathway due to activation of one or more of indoleamine
2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase enzymes. The compounds of the invention are useful
in the treatement of immune autoimmunity. In one aspect,
autoimmunity are associated with one or more of indoleamine
2,3-dioxygenase-1 or indoleamine 2,3-disoxygenase-2 or tryptophan
2,3-dioxygenase enzymes.
[0305] The term "regulation" or variations thereof as used herein
refers to the ability of a compound of formula (I) to inhibit one
or more components of a biological pathway. In one embodiment,
"regulation" refers to a decrease in plasma and/or tissue
concentrations of kynurenine. In another embodiment, "regulation"
refers to a decrease in plasma and/or tissue concentrations of
kynurenine/tryptophan (kyn/trp) ratio. In yet another embodiment,
"regulation" referes to an increase in plasma and/or tissue
concentations of tryptophan. In still another embodiment,
"regulation" refers to (i) a decrease in concentrations of
kynurenine and/or kyn/trp ratio, and/or (ii) increase in tryptophan
concentration in an in vitro assay, for example, using cell culture
system.
[0306] In another embodiment, "regulation" refers to inhibition of
indoleamine 2,3-dioxygenase-1 activity. In yet another embodiment,
"regulation" refers to inhibition of indoleamine 2,3-dioxygenase-2
activity. In another embodiment, "regulation" refers to inhibition
of tryptophan 2,3-dioxygenase activity. In a further embodiment,
"regulation" refers to dual inhibition of indoleamine
2,3-dioxygenase-1 and tryptophan 2,3-dioxygenase activity. In a yet
further embodiment, "regulation" refers to dual inhibition of
indoleamine 2,3-dioxygenase-2 and tryptophan 2,3-dioxygenase
activity. In a still further embodiment, "regulation" refers to
dual inhibition of indoleamine 2,3-dioxygenase-1 and indoleamine
2,3-dioxygenase-2 activity. In a still further embodiment,
"regulation" refers to triple inhibition of indoleamine
2,3-dioxygenase-1, indoleamine 2,3-dioxygenase-2 and tryptophan
2,3-dioxygenase activity.
[0307] The utility of the compounds can be illustrated, for
example, by their activity in in vitro and in vivo assays known in
the art and as described herein. The compounds of formula (I) or
metabolites thereof, or a pharmaceutically accetable salt or
prodrug thereof exhibit indoleamine 2,3-dioxygense-1 and/or
indoleamine 2,3-disoxygenase-2 and/or tryptophan 2,3-dioxygenase
inhibitory activity, and decrease the production of kynurenine
pathway metabolites. Accordingly, compounds of the invention can be
used as therapeutic agents for the treatment of a disease,
disorder, or condition directly or indirectly related to or
associated with kynurenine pathway metabolites and/or one or more
of indoleamine 2,3-dioxygenase-1, indoleamine 2,3-dioxygenase-2 and
tryptophan 2,3-dioxygenase enzymes.
[0308] As used herein, "disease," "disorder" and "condition" are
used interchangeably, to indicate an abnormal state in a subject.
Kynurenine pathway associated disease is a disease that can be
treated, prevented, ameliorated or cured by reducing kynurenine
pathway metabolite levels or increasing tryptophan levels or both.
IDO1-, IDO2-, and/or TDO-associated disease can be any disease that
can be treated, prevented, ameliorated or cured by regulating
enzyme expression and/or activity. The association may be direct or
indirect. Accordingly, the compounds described herein are useful
for treating diseases associated directly or indirectly with IDO1,
IDO2 or TDO or any combination these enzymes, or with kynurenine
pathway.
[0309] In one embodiment, such a disease is associated with
abnormal cellular proliferation. The term "abnormal cellular
proliferation" refers to the uncontrolled growth of cells which are
naturally present in a mammalian body. In one embodiment, a disease
which is characterized by abnormal cellular proliferation is
cancer, including, without limitation, squamous cell cancer (e.g.,
epithelial squamous cell cancer), cancer of the peritoneum,
prostate, head, neck, eye, mouth, throat, esophagus, bronchus,
larynx, pharynx, thyroid cancer, chest, bone, lung including
small-cell lung cancer ("SCLC"), non-small cell lung cancer
("NSCLC"), adenocarcinoma of the lung and squamous carcinoma of the
lung, colon, rectum, gastric or stomach including gastrointestinal
cancer, bladder, uterus, cervix, breast, ovaries, uterus including
endometrial or uterine carcinoma, vagina, vulval cancer, testicles,
penile carcinoma, anal carcinoma, skin, thyroid, blood, lymph
nodes, kidney or renal, liver including hepatocellular cancer,
hepatic carcinoma, and hepatoma, intestines, salivary gland
carcinoma, pancreas, brain including glioblastoma, central nervous
system, adrenal gland, skin or leukemia. Accordingly, the use of
compounds for treating cancer is provided. One of skill in the art
would understand that there is an established link between
decreased kynurenine levels and anti-tumor activity in the clinical
setting.
[0310] As described herein, a therapeutically effective amount of a
compound when used for the treatment of cancer is an amount which
may reduce the number of cancer cells, reduce tumor size, inhibit
metastasis, inhibit or reduce tumor growth, reduce tumor
resistance, reduce tumor evasion, and/or ameliorate one or more of
the symptoms of the cancer. For cancer therapy, efficacy can be
measured for example, by assessing the time to disease progression
and/or determining the response rate.
[0311] In one embodiment, the condition is immunosuppression. The
terms "immune suppression" or "immunosuppression" used throughout
the present disclosure refers to suppression of the body's immune
system and its ability to fight infections and other diseases. The
suppression of the immune system may be partial or complete.
Immunosuppression may result from certain diseases, for example
without limitation, abnormal cell proliferation or cancer, acute
and chronic infections including but not limited to bacterial
infections such as tuberculosis, viral infections such as AIDS,
HIV, HCV, HPV infection and parasitic infections such as malaria
and Leishmaniasis. In addition, immunosuppression may result from
disease treatment, for example, from treatment with anticancer
drugs such as tetracycline or its analogs. The compounds as
described herein are useful for treating immunosuppression. More
specifically, the compounds can be utilized in order to reduce
immune suppression associated with abnormal cell growth in which
one or more of indoleamine 2,3-disoxygenase-1 or indoleamine
2,3-disoxygenase-2 or tryptophan 2,3-disoxygenase plays a role.
Thus, the compounds are effective in the treatment of diseases,
disorders or conditions such as cancer, associated with increased
kynurenine levels due to the actions of indoleamine
2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2 and/or
tryptophan 2,3-dioxygenase.
[0312] Where immunosuppression is desired for treatment of a
condition or disease or for a procedure such as preparation for
bone marrow or other organ transplantation, immunosuppression is
generally induced with drugs to prevent rejection of the donor
tissue. Examples include without limitation allogeneic
hematopoietic stem cell transplantation (HSCT), graft-versus-host
disease (GvHD), organ transplant etc. The present invention also
provides for use of the compounds of the invention and
pharmaceutical compositions for inducing faster recovery from
immunosuppression after bone marrow treatment or other organ
transplantation to fight infection post-procedure. The timing for
the use of the compounds will be determined by a healthcare
professional or a physician. The compounds may also be used as
adjuvants after bone marrow transplantation or peripheral blood
stem cells transplantation and in immunotherapy by adoptive
transfer. Accordingly, the use of compounds as described herein for
treating conditions with immune suppression is provided.
[0313] In another embodiment, the disease is infectious disease. In
one embodiment, infectious disease is a bacterial infection.
Examples of bacterial infections treatable include but are not
limited to Mycobacteria infection and Streptococcus pyrogens
infection. Particular intracellular bacterial infections may be
selected from the group consisting of Mycobacterium leprae,
Mycobacterium tuberculosis, Listeria monocytogens and Toxoplasma
gondii. Accordingly, use of compounds for the treatment of
bacterial infection is provided herein.
[0314] In a further embodiment, the disease is a viral infection
(for example HIV, HPV, or HCV infection). Examples of viral
infections that may be treated using compounds of the invention
include but not limited to are Human immunodeficiency (HIV)/AIDS
virus, human parainfluenza virus, human papilloma virus, Hepatitis
C, Hepatitis B, influenza, SARS, cytomegalovirus, viral hemorrhagic
fevers (Ebola, Marburg, Lassa and yellow fever virus), polio virus,
Epstein Barr virus, Varicella zoster virus and Coxsackie virus. In
another embodiment, the viral infection is HCV infection. In
another embodiment, the viral infection is HIV infection.
Accordingly, the use of compounds as described herein for treating
viral infections, such as HIV, HPV, or HCV infection, is
provided.
[0315] In still another embodiment, the disease is a parasitic
disease. Examples of parasitic diseases that may be treated using
compounds of the invention include but not limited to are
Leishmaniasis and Malaria. The compounds of the invention and
pharmaceutical compositions are useful in the treatment against
parasites that include but are not limited to Leishmania donovani,
Leishmania tropica, Leishmania major, Leishmania aethiopica,
Leishmania maxicana, Plasmodium falciparum, Plasmodium vivax,
Plasmodium ovale and Plasmodium malariae. Accordingly, the use of
compounds as described herein for treating parasitic disease such
as Leishmaniasis and malaria is provided.
[0316] In a still further embodiment, the disease is an immune
mediated disorder (for example a B-cell mediated disorder or a
macrophage-mediated disorder, or a T-cell mediated immune disease).
IDO1, for example, is induced by pro-inflammatory cytokines such as
Interferon gamma and to a lesser extent by TNF-alpha, IL-1,
IFN-alpa and beta. IDO2 is a critical mediator of inflammatory
pathogenesis and autoreactive responses in autoimmune arthritis.
Accordingly, in one embodiment, the disease is an inflammatory
disease. Examples of inflammatory disorders or conditions include,
without limitation, arthritis, pulomonary disease, allergic airway
disease, asthma, cardiovascular and neurovascular diseases such as
Atherosclerosis, coronary artery disease, peripheral artery
disease, ischemia, Alzheimer's disease, and stroke, Irritable bowel
syndrome, Crohn's disease, and pelvic inflammatory disorder. In one
embodiment, the disease is arthritis. In another embodiment,
arthritis is selected from a group consisting of osteoarthritis,
rheumatoid arthritis, juvenile arthritis, ankylosing spondylitis,
gout, and psoriatic arthritis. One of skill in the art will
recognize that inhibition of IDO 1 activity reduces circulating
T-rcgs and effector T-cell apoptosis in subjects with inflammatory
disorders and promotes immune tolerance. Thus, the compounds
described herein are useful for treating or regulating inflammatory
disorders associated with one or more of IDO1 or IDO2 or TDO
enzymes. The compounds are used for treating or regulating
inflammatory disorders associated with kynurenine pathway
metabolites. In one embodiment, the compounds are useful for
treating IDO 1 associated immune-mediated diseases. In another
embodiment, the compounds are useful for treating IDO2 associated
immune-mediated disorders. In another embodiment, the compounds are
useful for treating or regulating IDO2 associated inflammatory
diseases. In another embodiment, the compounds are useful for
treating TDO associated immune-mediated disorders. In yet another
embodiment, the compounds are useful for treating or regulating one
or more of (i) T-cell mediated immune disorders, or (ii) B-cell
mediated immune disorders, or (iii) macrophage- or mast-cell
mediated immune disorders. Accordingly, the use of compounds as
described herein for treating immune-mediated disorders, including
inflammatory diseases, is provided. In one embodiment, the
compounds are useful for treating or regulating osteoarthritis,
rheumatoid arthritis, juvenile arthritis, anklylosing spondylitis,
and psoriatic arthritis. In another embodiment, compounds are
useful for treating or regulating Alzheimer's disease.
[0317] IDO2 has been shown to inhibit the production of autoimmune
antibodies in mouse model of autoimmune arthritis (Merlo et al. J.
Immunol. (2014), 192(5), 2082-2090). Accordingly in one embodiment,
the disease is autoimmune disorder. Examples of autoimmune
disorders that may be treated using the compounds of the invention
include, without limitation, multiple sclerosis, ulcerative
colitis, rheumatoid arthritis, asthma, psoriasis, inflammatory
bowel disease, primary sclerosing cholangitis, Hashimoto's
thyroiditis, Sjogren's syndrome, systemic lupus erythematosus,
antiphospholipid syndrome--primary and secondary, primary biliary
cirrhosis, autoimmune hepatitis, encephalomyelitis, Graves'
disease, autoimmune retinopathy--also called recoverin-associated
retinopathy, scleroderma, autoimmune thrombocytopenic purpura,
Addison's disease, celiac disease sprue, dermatomyositis, chronic
inflammatory demyelinating polyradiculoneuropathy, acute
inflammatory demyelinating polyneuropathy, Isaacs' syndrome,
Moersch-Woltmann syndrome, Lambert-Eaton myasthenic syndrome, and
myasthenia gravis. In one embodiment, the disease is rheumatoid
arthritis. In another embodiment, the disease is multiple
sclerosis. In yet another embodiment, the disease is lupus
erythromatosis. Accordingly, the use of compounds as described
herein for treating autoimmune disease is provided. In one aspect,
the use of compounds as described herein for treating multiple
sclerosis is provided. In another aspect, the use of compounds as
described herein for treating rheumatoid arthristis is
provided.
[0318] IDO1 and TDO are expressed in the brain and are highly
expressed in brain tumors. IDO1 activity in brain tumors negatively
impacts survival (Wainright, et al. Clin Cancer Res. 2012 Nov. 15;
18(22):6110-21). Brain IDO1 contributes to the comorbidity of pain
and depression (Kim et al. J Clin Invest. 2012; 122(8):2940-2954).
Accordingly in another embodiment, the disease is a disease of the
nervous system. Examples of diseases of central and peripheral
nervous system that may be treated using the compounds of the
invention include, without limitation, brain tumors such as glioma,
giobastoma, neuroma, neuroinflammatory and neurodegenerative
diseases such as Alzheimer's disease, Huntington's disease,
multiple sclerosis, amyotrophic lateral sclerosis, and Parkinson's
disease, lyme neuroborreliosis, late lyme encephalopathy,
Tourette's syndrome, systemic sclerosis, Guillain-Barr syndrome,
muscular dystrophy, acute disseminated encephalomyelitis, and optic
neuritis, transverse myelitis, neuromyelitis optica. Examples of
diseases also include without limitation neuropsychiatric diseases,
including mood disorders and sleep disorders. In another
embodiment, the disease is depression. In another embodiment, the
disease is schizophrenia. In yet another embodiment, the sleep
disorder is insomnia. In still another embodiment, the sleep
disorder is sleep apnea. Accordingly, the use of compounds as
described herein for treating diseases of the nervous system is
provided. In one aspect, the use of compounds as described herein
for treating multiple sclerosis is provided. In one aspect, the use
of compounds as described herein for treating Alzheimer's disease
is provided. In another aspect, the use of compounds as described
herein for treating depression is provided. In yet another aspect,
the use of compounds as described herein for treating schizophrenia
or sleep disorder is provided.
[0319] In another aspect, a method for regulating a kynurenine
pathway is provided and includes administering a compound a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need thereof In
one aspect, the disease may be any disease treatable by
adminstering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof is provided. In
another aspect, a method for treating a disease treatable by
inhibiting a kynurenine pathway is provided and includes
administering a compound, a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to a subject in
need thereof.
[0320] In another aspect, a method of regulating any one or more of
any one or more of indoleamine 2,3-dioxygenase-1 or an indoleamine
2,3-dioxygenase-2 or a tryptophan 2,3-dioxygenase enzymes is
provided and includes administering a compound of formula (I) (IV),
a metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof
[0321] In another aspect, the regulating is inhibiting the
kynurenine pathway or one or more of the enzymes.
[0322] In still a further aspect, a method of regulating the
kynurenine pathway by inhibiting indoleamine 2,3-dioxygenase-1
and/or tryptophan 2,3-dioxygenase is provided and includes
administering a compound as described herein to a subject in need
thereof.
[0323] In still a further aspect, a method of regulating the
kynurenine pathway by inhibiting indoleamine 2,3-dioxygenase-2
and/or tryptophan 2,3-dioxygenase is provided and includes
administering a compound as described herein to a subject in need
thereof.
[0324] In still a further aspect, a method of regulating the
kynurenine pathway by inhibiting indoleamine 2,3-dioxygenase-1
and/or indoleamine 2,3-dioxygenase-2 is provided and includes
administering a compound as described herein to a subject in need
thereof.
[0325] In one aspect, a method of reducing kynurenine pathway
metabolites is provided and includes administering a compound or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof
[0326] In another aspect, a method of altering tryptophan levels in
a subject and includes administering a compound of formula (I) or a
metabolite thereof, or a pharmaecuticallly acceptable salt or
prodrug thereof described herein is provided. In one aspect, the
tryptophan levels are increased. In another aspect,
kynurenine/tryptophan ratio is decreased.
[0327] In yet another aspect, a method for increasing tryptophan
levels is provided by inhibiting one or more of indoleamine
2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase is provided and includes administering a compound
as described herein to a subject in need thereof.
[0328] In one aspect, a method of treating a disease associated
with or resulting from dysregulation of a kynurenine pathway is
provided and includes administering a compound of formula ((I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof.
[0329] In another aspect, a method for treating a disease caused by
the dysregulation of the kynurenine pathway by inhibiting
indoleamine 2,3-dioxygenase-1 and/or indoleamine 2,3-dioxygenase-2
and/or tryptophan 2,3-dioxygenase is provided and includes
administering a compound or prodrug thereof described herein to a
subject in need thereof
[0330] In yet another aspect, a method for treating a disease
caused by activation of indoleamine 2,3-dioxygenase-1 or tryptophan
2,3-dioxygenase or both enzymes is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or propdrug thereof as described
herein to a subject in need thereof.
[0331] In yet another aspect, a method for treating a disease
caused by activation of indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase enzymes or both is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or propdrug thereof as described
herein to a subject in need thereof.
[0332] In yet another aspect, a method for treating a disease
caused by activation of indoleamine 2,3-dioxygenase-1 or
indoleamine 2,3-dioxygenase-2 or both is provided and includes
administering a compound of formula (I) or a metabolite thereof, or
a pharmaceutically acceptable salt or propdrug thereof as described
herein to a subject in need thereof.
[0333] In another aspect, a method of inhbiting activation of one
or more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes is provided
and includes administering a compound of formula (I) or a
metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof as described herein to a subject in need
thereof.
[0334] In another aspect, a method for treating a disease
associated with any one or more of indoleamine 2,3-dioxygenase-1 or
indoleamine 2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes
is provided and includes administering a compound of formula (I) or
a metabolite of the compound, or a pharmaceutically acceptable salt
or prodrug thereof described herein to a subject in need
thereof.
[0335] In yet a further aspect, a method is provided for treating a
disease characterized by abnormal immune suppression, for example,
increased immune suppression resulting from dysregulation of the
kynurenine pathway and includes administering a compound of formula
(I) or a metabolite thereof, or pharmaceutically acceptable or
prodrug thereof described herein to a subject in need thereof.
[0336] In a further aspect, a method for regulating a disease
characterized by abnormal immune suppression resulting from a
dysregulated kynurenine due to activation one or more of
indoleamine 2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or
tryptophan 2,3-dioxygenase enzyme is provided and includes
administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof as described
herein to a subject in need thereof.
[0337] In one aspect, a method of treating immune suppression is
provided and includes administering a compound or a metabolite
thereof or a pharmaceutically acceptable salt or prodrug thereof as
described herein to a subject in need thereof. In another aspect,
the immune suppression is associated with increased kynurenine
metabolite levels or enzymatic activity of one or more of
indoleamine 2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or
tryptophan 2,3-dioxygenase enzymes. In yet another aspect, a method
is provided for treating immune suppression associated with one or
more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes and
includes administering a compound or a metabolite thereof, or
pharmaceutically acceptable salt or prodrug thereof to a subject in
need thereof. Thus, in an aspect, compounds of the invention for
use in the treatment of immunosuppression associated with one or
more of indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase enzymes are
provided.
[0338] In yet another aspect, a method for treating immune
suppression through inhibiting enzymatic activity of indoleamine
2,3-dioxygenase and/or tryptophan 2,3-dioxygenase is provided and
includes administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a subject in need thereof.
[0339] In yet another aspect, a method of reducing or eliminating
an immune mediated disorder is provided and includes administering
a compound or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof described herein to a
patient.
[0340] In yet another aspect, a method of inhibiting an autoimmune
reaction or autoimmune antibody production in a subject is provided
and includes administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof as described to
a subject in need thereof. In another aspect, a method of
inhibiting autoimmune reaction or autoimmune antibody production is
provided wherein (i) indoleamine 2,3-dioxygenase-2 is inhibited or
(ii) kynurenine metabolites are reduced, and includes administering
a compound or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof as described to a subject in
need thereof. In one aspect, the foregoing reduction in autoimmune
reaction or autoimmune antibody production is associated with
inflammatory diseases, cancer or autoimmine disorders.
[0341] In one aspect, diseases that can be treated using compounds
of the invention comprise cancer, bacterial infection, viral
infection, parasitic infection, immune-mediated disorder,
autoimmune disorder, inflammatory disease, central nervous system
disease, peripheral nervous system disease, neurodegenerative
disease, mood disorder, sleep disorder, cerebrovascular disease,
peripheral artery disease, or cardiovascular disease. In another
aspect, all foregoing methods comprise administration of one or
more additional medication or therapeutic agent or therapy. In one
aspect, the therapeutic agent is a chemotherapeutic agent selected
from a group further comprising a cancer vaccine, a targeted drug,
a targeted antibody, an antibody fragment, an antimetabolite, an
antincoplastic, an antifolatc, a toxin, an alkylating agent, a DNA
strand breaking agent, a DNA minor groove binding agent, a
pyrimidine analog, a purine analog, a ribonucleotide reductase
inhibitor, a tubulin interactive agent, an anti-hormonal agent, an
immunomoldulator, an anti-adrenal agent, a cytokine, a radiation
therapy, a cell therapy, cell depletion therapy such as B-cell
depletion therapy, or a hormone therapy.
[0342] In another aspect, a method of treating depression,
Alzheimer's disease, dementia, multiple sclerosis, schizophrenia,
HIV inection, malaria, rheumatoid arthritis, or insomnia is
provided and includes administering a compound or a metabolite
thereof, or a pharmaceutically acceptable salt or prodrug thereof
described herein to a patient.
[0343] In one aspect, the disease is cancer. In another aspect,
cancer disease is a cancer of squamous cell, peritoneum, prostate,
head, neck, eye, mouth, throat, esophagus, bronchus, larynx,
pharynx, thyroid cancer, chest, bone, lungs, colon, rectum,
stomach, urinary bladder, gall bladder, uterus, cervix, breast,
ovaries, uterus, vagina, vulva, testicles, penis, anus, skin,
thyroid, blood, lymph nodes, kidney, liver, intestines, salivary
gland, pancreas, brain, spine, adrenal gland, skin or leukemia. In
another aspect, a method of treating tumor resistance is provided
comprising administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a patient.
[0344] In another aspect, the viral infection is HIV infection. In
another aspect, parasite infection is malaria or Leishmaniasis.
[0345] In a further aspect, a method of treating a viral infection
is provided and includes administering a compound described herein
to a patient.
[0346] In another aspect, a method of treating depression is
provided and includes administering a compound described herein to
a patient.
[0347] In still another aspect, a method of treating schizophrenia
is provided and includes administering a compound described herein
to a patient.
[0348] In still another aspect, a method of treating Alzheimer's
disease is provided and includes administering a compound described
herein to a patient.
[0349] In one embodiment, methods for regulating the kynurenine
pathway by inhibiting indoleamine 2,3-dioxygenase-1 or indoleamine
2,3-dioxygenase-2 or tryptophan 2,3-dioxygenase are provided and
include administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or a prodrug thereof to a patient
in need thereof.
[0350] In another embodiment, methods for treating a disease
characterized by abnormal immune suppression resulting from a
dysregulated kynurenine pathway due to activation of indoleamine
2,3-dioxygenase-1 or indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase arc provided and include administering of a
compound or a metabolite thereof, or a pharmaceutically acceptable
salt or a prodrug thereof to a patient in need thereof.
[0351] In another embodiment, methods for treating a disease
characterized by an abnormal immune suppression resulting from a
dysregulated kynurenine pathway due to activation of one or more of
indoleamine 2,3-dioxygenase-1 or or indoleamine 2,3-dioxygenase-2
or tryptophan 2,3-dioxygenase are provided and include
administering of a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or a prodrug thereof to a patient
in need thereof.
[0352] In yet another aspect, a method for diagnosing and treating
a disease associated with kynurenine pathway or any one or more of
indoleamine 2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2
or a tryptophan 2,3-dioxygenase enzymes in a subject is provided
and includes: (i) assaying a blood and/or tissue sample from a
subject; (ii) determining the subject's blood and/or tissue
tryptophan or Kynurenine concentration or both in the sample; (iii)
optionally determining the subject's Kynurnine/tryptophan ratio;
and (iv) administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof described
herein to a subject.
[0353] In still another aspect, a method of monitoring a disease
associated with kynurenine pathway or one or more of indoleamine
2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2 or a
tryptophan 2,3-dioxygenase enzymes in a subject is provided and
includes (i) dosing a subject having a disease associated with
kynurenine pathway with a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof, (ii) analyzing
a blood or tissue sample or both at one or more time points or
continuously during a treatment regimen, (iii) determining a
tryptophan and a kynurenine concentration in the blood or the
tissue sample or both, (iv) optionally determining the subject's
kynurnine/tryptophan ratio, and (v) adjusting the treatment regimen
or dosage of the compound.
[0354] In a further aspect, a method for diagnosing and treating a
disease associated with kynurenine pathway or any one or more of
indoleamine 2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2
or a tryptophan 2,3-dioxygenase enzymes in a patient is provided
and includes (i) analyzing a patient sample for the presence or
absence of altered kynurenin/tryptophan ratio, wherein the patient
is diagnosed with a disease associated with kynurenine pathway if
altered kynurenine/tryptophan ratio is detected and (ii)
administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug to the diagnosed
patient.
[0355] In still a further aspect, a method for treating a disease
associated with kynurenine pathway or one or more of an indoleamine
2,3-dioxygenase-1 or an indoleamine 2,3-dioxygenase-2 or a
tryptophan 2,3-dioxygenase enzyme in a patient and includes (i)
requesting a test providing the results of an analysis to determine
whether the patient's kynurninc levels arc altered, and (ii)
administering a compound or a metabolite thereof, or a
pharmaceutically acceptable salt or prodrug thereof to the patient
if the patient's kynurenine levels are altered.
[0356] In another aspect, provided herein are compounds for use in
a disease associated with kynurenine pathway in a subject
comprising: assaying a blood sample from a patient; determining if
a patient has high blood and/or tissue kynurenine levels; and
administering a compound as described herein to the patient if
blood and/or tissue kynurenine levels are high.
[0357] In one aspect, provided herein are compounds for use for
treating disease associated with kynurenine pathway in a patient
comprising: assaying a blood sample from a patient; determining if
a patient has low blood and/or tissue tryptophan levels; and
administering a compound as described herein to the patient if
blood and/or tissue tryptophan levels are low.
[0358] In one aspect, provided herein are compounds for use for
treating a disease associated with one or more indoleamine
2,3-dioxygenase-1 or or indoleamine 2,3-dioxygenase-2 or tryptophan
2,3-dioxygenase in a patient comprising: assaying a blood sample
from a patient; determining if a patient has high blood and/or
tissue kynurenine levels or low blood and/or tissue tryptophan
levels; and administering an amount of a compound as described
herein to the patient if blood and/or tissue kynurenine levels are
high or blood and/or tissue tryptophan levels are low.
[0359] In yet another aspect, a use of foregoing methods is
provided wherein the disease is cancer, bacterial infection, viral
infection, parasitic infection, immune-mediated disorder,
autoimmune disorder, inflammatory disease, central nervous system
disease, peripheral nervous system disease, neurodegenerative
disease, mood disorder, sleep disorder, cerebrovascular disease,
peripheral artery disease, or cardiovascular disease.
[0360] In one aspect, provided herein are compounds for use in
treating immune suppression in a patient comprising: assaying a
blood sample from a patient; determining if a patient has high
blood and/or tissue kynurenine levels, and/or low blood and/or
tissue tryptophan levels; and administering a compound as described
herein to the patient if blood and/or tissue kynurenine levels are
high, and/or blood and/or tissue tryptophan levels are low.
[0361] In another aspect, a method for treating a disease
associated with kynurenine pathway in a mammal is provided, said
method comprising: assaying a blood and/or tissue sample from a
mammal; determining the mammal's blood and/or tissue tryptophan and
kynurenine concentrations; optionally determining the mammal's
kynurnine/tryptophan ratio; and administering an amount of a
compound described herein.
[0362] In one aspect, a method for treating a disease associated
with kynurenine pathway in a mammal is provided, said method
comprising: assaying blood and/or tissue sample from a subject;
determining the patient's IDO1 and/or IDO2 and/or TDO expression
and/or activity in tissue sample; and administering an amount of a
compound as described herein.
[0363] In yet another aspect, a method of monitoring or tracking a
disease associated with kynurenine pathway in a mammal is provided,
said method comprising: dosing a mammal having a disease associated
with kynurenine pathway with a compound in combination with one or
more therapeutic agent; analyzing a blood and/or tissue samples at
one or more time points or continuously during the treatment
regimen; determining the mammal's blood and/or tissue tryptophan
and kynurenine concentrations and/or determining the
kynurenine/tryptophan ratio; adjusting the treatment regimen or
dosage of the compound or the second therapeutic agent.
[0364] In another aspect, a method of monitoring or tracking a
disease associated with kynurenine pathway in a mammal is provided,
said method comprising: dosing a mammal having a disease associated
with kynurenine pathway with a compound as described herein;
analyzing a blood and/or tissue sample of the mammal at one or more
time points or continuously during the treatment regimen;
determining the blood and/or tissue dosed compound's metabolite
concentration; and adjusting the treatment regimen or therapeutic
dosage.
[0365] The compounds of the invention may used in combination with
one or more therapeutic agents as described herein. The compounds
of the invention are thus useful in the treatment and monitoring
the progression of disease associated with kynurenine pathway.
[0366] In yet another aspect, a method for diagnosing and treating
a disease associated with kynurenine pathway in a patient
comprising: analyzing a patient sample for the presence or absence
of altered kynurenine and/or tryptophan and/or kynuren i n/tryptoph
an ratio, wherein the patient is diagnosed with a disease
associated with kynurenine pathway if altered kynurenine and/or
tryptophan and/or kynurenine/ratio is detected, and administering
therapeutically effective amount of a compound to the diagnosed
patient.
[0367] In yet another aspect, a method for treating a disease
associated with kynurenine pathway in a patient comprising:
requesting a test providing the results of an analysis to determine
whether the patient's kynurnine and/or tryptophan levels and/or
kynurenine/tryptophan ratio are altered and administering
therapeutically effectively amount of a compound to the patient if
the patient's kynurenine and/or tryptophan levels and/or
kynurenine/tryptophan ratio are altered.
[0368] In a further aspect, a method for diagnosing and treating
cancer associated with IDO1 or IDO2 or TDO in a subject, wherein
the cancer is characterized by increased expression and/or activity
of one or more of IDO1 or IDO2 or TDO biomarkers, said method
comprising: i) obtaining a biological sample from the subject; ii)
applying a monoclonal antibody specific for IDO1 or IDO2 or TDO to
the sample, wherein presence of IDO 1 or IDO2 or TDO creates an
antibody-IDO1 or an antibody-IDO2 or antibody-TDO biomarker
complex; iii) applying a detection agent that detects the
antibody-biomarker complex; iv) diagnosing cancer associated with
one or more of IDO1 or IDO2 or TDO wherein the detection agent of
step iii) is detected; and v) administering to the subject a
compound or a metabolite thereof, or a pharmaceutically acceptable
salt or prodrug thereof.
[0369] In yet another aspect, a method for treating a disease
associated with kynurenine pathway in a patient comprising:
requesting a test providing a result of an analysis to determine
whether the patient's IDO 1 and/or IDO2 and/or TDO expression
and/or activity are increased and administering therapeutically
effectively amount of a compound to the patient if the patient's
IDO1 and/or IDO2 and/or TDO expression and/or activity are
increased. Measurement of tryptophan, kynurenine pathway
metabolites such as kynurenine, and/or metabolites of the compounds
of the invention can be done vivo or in vitro using methods known
in the art, including without limitation, HPLC, LC/MS/MS,
Fluorescence, ELISA, RIA techniques and continuous monitoring
sensor technologies using devices placed over the skin or eye, or
inserted into the skin or tissue. Measurement of IDO1 and/or IDO2
and/or TDO expression and/or activity can be done as a non-limiting
example using in vitro assays known in the art such as PCR, ELISA,
enzymatic assays and as described herein.
[0370] The present invention, thus, also provides compounds and
methods of assaying the activity IDO1 and/IDO2 and/or TDO in a
cell-free system or in a system containing cells expressing IDO1
and/or IDO2 and/or TDO (such as a cell culture system, tissue,
living organism such as a mammal, or in plasma or serum) comprising
contacting test sample with a compound of the invention and
measuring the inhibition of tryptophan degradation or catabolism
and the reduction in kynurenine pathway metabolite levels as
compared to a control treated samples.
Combination Therapy
[0371] It is within the scope of the invention to combine one or
more compounds of formula (I)-(IV) with one or more metabolite of
the compounds of the formula (I)-(IV) and/or with one or more
prodrug of the compounds of formula (I). In addition to the
components described above for use in compositions and the compound
of formula (I) or a metabolite thereof, or pharmaceutically
acceptable salts or prodrug thereof, the compositions may contain
one or more medications or therapeutic agents which are used to
treat solid tumors and other diseases described herein.
Therapeutically effective amounts of the additional medication(s)
or therapeutic agents are well known to those skilled in the art.
However, it is well within the attending physician or healthare
procssional to determine the amount of other medication to be
delivered.
Combination therapy for the treatment of Cancer
[0372] In one embodiment, the additional medication is a
chemotherapeutic. Examples of chemotherapeutics include those
recited in the "Physician's Desk Reference", 64.sup.th Edition,
Thomson Reuters, 2010, which is hereby incorporated by reference.
Methods for the safe and effective administration of most of these
chemotherapeutic agents are known to those skilled in the art. In
addition, their administration is described in the standard
literature. For example, the administration of many
chemotherapeutic agents is described in "Physician's Desk
Reference" (PDR, e.g., 2010 edition, PDR Network, Montvale, N.J.),
the disclosure of which is also incorporated by reference in its
entirety. In one aspect, the chemotherapeutic is doxorubicin,
paclitaxel or derivative thereof, 5-FU, and carboplatin or a
derivative thereof.
[0373] Suitable antineoplastic chemotherapeutic agents that can be
dosed in combination with the compounds of invention can include,
for example without limitation, alkylating agents (including
without limitation, nitrogen mustards, ethylenimine derivatives,
alkyl sulfonates, nitrosoureas, and triazine), uracil mustard,
cyclophosphamide (Cytoxan.TM.), chlormethine, ifosfamide, melphala,
chlorambucil, pipobroman, triethylene melamine,
triethylenethiophosphoramine, busulfan, carmustine, lomustine,
streptozocin, dacarbazine, temozoloide, and combinations
thereof.
[0374] Other chemotherapeutic or anti-cancer agents include, for
example without limitation, antimetabolites (including without
limitation, folic acid antagonists or antifolates, pyrimidine
analogs, purine analogs, and adenosine deaminase inhibitors) such
as methotrexate, fluorouracil, gemcitabine, and combinations
thereof. Suitable chemotherapeutic or anti-cancer agents further
include certain natural products and their derivatives, for example
without limitation, vinca alkaloids, anti-tumor antibiotics,
enzymes, lymphokines, and epipodohyllotoxins) such as vinblastine,
doxorubicin, vincristine, vindesine, bleomycin, dactinomycin,
daunorubicin, epirubicin, idarubicin, ara-C, paclitaxel
(TAXOL.TM.), deoxycoformycin, mitomycin-C, mithramycin,
L-asparagine, interferons (particularly IFN-a), etoposide, and
teniposide and combinations thereof.
[0375] The compounds may be used to augment the effects of
therapeutic vaccination against various tumors. When the compounds
are used in combination, then at least one additional therapeutic
agent may be a vaccine. The vaccine may also be a tumor vaccine or
a melanoma vaccine. Preferably, the tumor vaccine comprises
genetically modified tumors cells or genetically modified tumors
cell lines. In such cases, preferably the genetically modified
tumors cells or genetically modified cell lines has been
transfected to express granulocyte-macrophage stimulating factor
(GM-CSF). Alternatively, the vaccine may comprise one or more
immunogenic peptides, preferably immunogenic peptides of cancer
testis antigen (CTAgs). Such CTAgs and immunogenic peptides thereof
are well known in the art. CTAgs protein include MAGE, BAGE, GAGE,
SSX, NY-ESO-1, LAGE, SCP, CTSP, CT7, CT8, CT9, CT10, CT11, SAGE,
OY-TES-1, NY-SAR-35 and NY-BR-1. Several MAGe proteins are known,
including MAGE-A I , A3, A4, A5, A6, A8, A10,A12, B1, B2, B2, B4,
C1, C2 and C3 proteins. Several SSX proteins exist, including SSX1
and SSX2, SSX3 and SSX5. Vaccine may comprise one of more DNA
vaccines and recombinant viruses. Further the tumor vaccine may
comprise dendritic cells. In another embodiment, the additional
medication is a cancer vaccine. In an aspect, the cancer vaccine is
a dendritic cell based vaccine. In one aspect, the cancer vaccine
is the Provenge.RTM. vaccine (Dendreon Corp).
[0376] The present invention also contemplates that compounds of
invention may be used in combination with other anti-cancer agents
such as antibody therapeutics. In a further embodiment, the
additional medication is a targeted antibody, i.e., an antibody
which targets a specific tumor type. The term "antibody" is used in
the broadest sense and specifically covers intact monoclonal
antibodies, polyclonal antibodies, chimeric antibodies, humanized
antibodies, multispecific antibodies (e.g. bispecific antibodies)
formed from at least two intact antibodies, and antibody fragments
so long as they exhibit the desired biological activity. The term
"Antibody fragments" comprise a portion of an intact antibody,
preferably the antigen binding or variable region of the intact
antibody. Examples of antibody fragments include Fab, Fab',
F(ab').sub.2, and Fv fragments; diabodies; linear antibodies
(Zapata et al. Protein Eng. 8(10):1057-1062, 1995); single-chain
antibody molecules; and multispecific antibodies formed from
antibody fragments. The targeted antibody may be a monoclonal or
polyclonal antibody and may be selected from those described in
Pasquetto et al., "Targeted Drug Delivery Using Immunoconjugates:
Principles and Applications", J. Immunother., 34(9):611-628
(November-December 2011), which is hereby incorporated by
reference.
[0377] In one aspect, the targeted antibody is one or more of
gemtuzumab (Mylotarg), alemtuzmab (CAMPATH.TM.), rituximab
(Rituxin, Mabthera), trastuzumab (Herceptin.TM.) nimotuxumab,
cetuximab (Erbitux), erlotinib (TARCEVA..RTM.., Genentech/OSI
Pharm.), bevacizumab (Avastin.TM.), pertuzumab (OMNITARG..RTM..,
rhuMab 2C4, Genentech), Brentuximab vedotin (Adcetris.TM.),
Ipilimumab (MDX-101 and also known as Yervoy), Ofatumumab
(Arzerra), Panitumumab (Vectibix), and Tositumomab (Bexxar), among
others. In another aspect, the targeted antibody is one or more of
alemtuzumab, apolizumab, aselizumab, atlizumab, bapineuzumab,
bevacizumab, bivatuzumab mertansine, cantuzumab mertansine,
cedelizumab, certolizumab pegol, cidfusituzumab, cidtuzumab,
daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab,
felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab
ozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab,
mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab,
nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab,
pascolizumab, pecfusituzumab, pectuzumab, pertuzumab, pexelizumab,
ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab,
rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab,
tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab,
tocilizumab, toralizumab, trastuzumab, tucotuzumab celmoleukin,
tucusituzumab, umavizumab, urtoxazumab, and visilizumab.ln another
embodiment, an additional medication includes antibodies to immune
co-stimulatory molecules including but not limited to CTLA-4, 4-1BB
and PD-1, antibodies to cytokines (including but not limited to
IL-10, TGF-beta, etc.), and chemokine receptors including but not
limited to CCR2, CCR4 etc., among others. In yet another
embodiment, the additional medication is a targeted drug. The term
"targeted drug" as used herein refers to a medication that blocks
cancer cell growth by interfering specific "targeted" molecules
which are required for tumor growth. See, Pasquetto cited above,
which is hereby incorporated by reference. In one aspect, the
targeted drug includes, without limitation, dasatnib, imatinib,
nilotinib, bosutnib, lcstaurtinib, ruxolitinib, crizotinib,
vandetabib, cabozantinib, afibercept, adipotide, denileukin
diftitox, everolimus, and temosirolimus, among others.
[0378] Other chemotherapeutic or anti-cancer agents include, for
example, cytotoxic agents such as platinum coordination agents (for
e.g., cisplastin, and carboplatin), antineoplastic enzymes,
topoisomerase inhibitors, biological response modifiers, growth
inhibitors, hematopoetic growth factors, immune modulators,
chemokines, cytokines (for example a granulocyte-macrophage colony
stimulating factor (GM-CSF) or FLT3-ligand), cell migration
blockers, and inhibitors of angiogenesis. Angiogenesis inhibitors
include, but are not limited to, angiostatin, endostatin,
thrombospondin, platelet factor 4, Cartilage-derived inhibitor
(CDI), retinoids, Interleukin-12, tissue inhibitor of
metalloproteinase 1, 2 and 3 (TIMP-1, TIMP-2, and TIMP-3) and
proteins that block the angiogenesis signaling cascade, such as
anti-VEGF (Vascular Endothelial Growth Factor) and IFN-alpha.
[0379] Alternatively, the compounds may be used to augment the
effects of radiation therapy, which may be delivered locally to the
tumor or to the whole body. In yet another embodiment, the
additional medication is hormonal therapy. The term "hormonal
therapy" as used herein refers to a medication that blocks cancer
cell growth by interfering with the activity of specific hormones
such as estrogen, testosterone, or dihydrotestosterone.
Combination Therapy for Viral Infections
[0380] When the compounds are used in combination, then at least
one additional therapeutic agent may be an anti-viral vaccine, for
example without limitation, anti-HIV-vaccine, anti-HCV vaccine, and
anti-HPV vaccine. Other suitable antiviral agents contemplated for
use in combination with the compounds of the present invention
comprise nucleoside and nucleotide reverse transcriptase inhibitors
(NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs),
protease inhibitors, and other anti-viral drugs. Examples of NRTIs
include zIDO1 vuine (AZT); didanosine (ddI); zalcitabine (ddC);
stavudine (d4T); lamivudine (3TC); abacavir (1592U89); adefovir
diplovixil [bis(POM)-PMEA]; lobucavir (BMS-180194); BCH-10652;
emitricitabine [(-)-FTC]; beta-L-FD4 (also known as beta-L-D4C and
beta-L-2',3'dicleoxy-5-fluoro-cytidene); DAPD, ((-)0b
eta-D-2,6,-diamino-purine dioxolane,); and Iodenosine (FddA).
Typical suitable NNRTIs include nevirpine (BI-RG-587); delaviradine
(BHAP, U-90152); efavirnz (DP-266); PNU-142721; AG-1549; MKC-442
(1-(ethoxy-methyl)-5-(1-methylethyl)-6-(phyenylmethyl)-2,4(1H,3H)-pyrimed-
inedione); and (+)-calanolide A(NSC-675451) and B. Typical suitable
protease inhibitors include saquinavir (Ro 31-8959); ritonavir
(ABT-538); indinavir (MK-639); nclfnavir (AG-1343); amprenavir
(141W94); lasinavir (BMS-234475); DMP-450; BMS-2322623; ABT-378;
and AG-1 549. Other antiviral agents include hydroxyurea,
ribavarin, IL-2, IL12, perntasufide and Yissum Project No. 11607.
In one aspect, the viral vaccine is an anti-HPV vaccine.
Combination Therapy for Bacteria and Parasitic Infections
[0381] When the compounds are used in combination, then at least
one additional therapeutic agent may be a anti-bacterial agent
(including without limitation, a vaccine against tuberculosis or
antibitoics) or an anti-parasitic therapeutic agent, such as
anti-parasitic vaccine, for example without limitation, a vaccine
against malaria. Other compounds that may be used in combination
with compounds of the invention include without limitation
chloroquinine, hydrocholoroquinine, ferroquinine, Artemisinin,
Atovaquone/Proguanil, Doxycycline, Mefloquine (Lariam), and
Primaquine. Anti-malarial vaccines include but are not limited to
RTS,S malaria vaccine, RTS,S-AS01 delayed fractional third dose,
Adenovirus (Ad35) vectored CS and RTS,S-AS01 in heterologous
prime-boost regimen, ChAd63/MVA ME-TRAP, ChAd63/MVA ME-TRAP+Matrix
M.TM., PfSPZ, polyepitope DNA EP1300, PfCelTOS FMP012, CSVAC,
ChAd63/MVA (CS; ME-TRAP), ChAd63/MVA (CS; ME-TRAP, AMA-1),
RTS,S/AS.sub.01B+ChAd63 and MVA encoding ME-TRAP, EBA175 RII,
FMP2.1/AS01B (AMA-13D7 E. coli expressed in AS01B adjuvant), GMZ2,
pfAMA1-DiCo, P27A, MSP3 [181-276] field, SE36, ChAd63 AMAl/MVA
AMA1, NMRC-M3V-Ad-PfCA, NMRC-M3V-D/Ad- PfCA Prime/Boost, PfPEBS,
ChAd63/AMA MVA/AMA1 +alhydrogel/CPG7909, ChAd63 MSP1/MVA MSP1,
Pfs25-EPA, Pfs25 VLP, ChAd63/MVA PvDBP. Anti-tuberculosis vaccines
include but are not limited to bacilli Calmetter-Guerin (BCG),
MVA85A, rBCG30, 72F fusion protein, ESAT6-Ag85b fusion protein, M.
tuberculosis antigens-antigens 85A, 85B and TB10.4. In one aspect,
the anti-parasitic vaccine is an anti-malaria vaccine.
Combination Therapy other Diseases
[0382] The present invention also relates to the use of compounds
of the invention to be used with one or more medicaments or
therapies to treat any disease that is treateable by use of the
compounds of the invention. For example, the compounds may be used
in combination with B-cell depletion therapy, targeted drug,
targeted antibody, vaccines, or other therapeutic agents for
inflammatory diseases, including without limitation, arthritis,
osteoarthrisitis, rheumatoid arthritis, juvenile arthritis, spinal
arthritis, psoriatic arthritis, ankylosing spondylitis,
fibromyaligia, gout, etc. Non-limiting examples of therapies used
in combination with compounds of the invention include steroids
(such as prednisone, dexamethasone), NSAIDS (such as Celebrex,
Meloxicam, ibuprofen), targeted antibodies or antibody fragments
such (Enbrel, Remicade/Infliximab, Humira, Rituxan/antiCD20),
antimetabolites and antifolates (such as Methotrexate) etc.
Compounds of the invention may also be used in combination for
treatment of autoimmune diseases (for example, Alzheimer's disease,
Huntington's disease, Parkinson's disease, multiple sclerosis etc),
cardiovascular diseases (for example, coronary artery disease,
peripheral artery disease, atherosclerosis and ischemia), and
kidney disease (such as end stage renal disease). It is intended
that where appropriate that combination therapies described
elsewhere in the specification is to be used in combination or the
treatment of the diseases listed herein.
[0383] The compounds or a metabolite thereof, or a pharmaceutically
acceptable salt or prodrug thereof, and/or other medication(s) or
therapeutic agent(s) may be administered in a single composition.
However, the present invention is not so limited. The compounds of
the invention may be administered sequentially, consecutively,
alternatingly, or in any manner a healthcare professional or a
physician deems appropriate. In other embodiments, the compounds or
a metabolite thereof, or a pharmaceutically acceptable salt or
prodrug thereof may be administered in one or more separate
formulations from other compounds of the invention or
chemotherapeutic agents, cancer vaccine, targeted drug, targeted
antibody, hormonal therapy or other agents as desired.
[0384] Reference are made in detail to certain embodiments of the
invention, examples of which are illustrated in the accompanying
structures and formulae. It is intended that any part of the
disclosure may be read in combination with any other part of the
disclosure, unless otherwise apparent from the text. While the
invention is described in conjunction with the enumerated
embodiments, it will be understood that they are not intended to
limit the invention to those embodiments. It is specifically
intended to cover all alternatives, modifications, and equivalents
which maybe included within the scope of the present invention as
defined by the claims. At various places in the present
specification, substituents of compounds of the invention may be
disclosed in groups. It is specifically intended that the invention
include each and every individual sub combination of the members of
such groups.
[0385] It is further appreciated that certain features of the
invention, which are, for clarity, described in the context of
separate embodiments, can also be provided in combination in a
single embodiment. Conversely various features of the invention
which are, for brevity, described in the context of a single
embodiment can also be provided separately or in any suitable
subcombination.
[0386] Furthermore, where the claims recite a composition, it is to
be understood that methods of using the composition for any of the
purposes disclosed herein are included, and methods of making the
composition according to any of the methods of making disclosed
herein or other methods known in the art arc included, unless
otherwise indicated or unless it would be evident to one of
ordinary skill in the art that a contradiction or inconsistency
would arise. In addition, the invention encompasses compositions
made according to any of the methods for preparing compounds and
compositions disclosed herein.
EXAMPLES
[0387] The following examples are illustrative only and are not
intended to limit the present invention. The following examples are
only meant to suggest a method of practicing the invention. One of
skill in the art will recognize that the chemical reactions
described may be readily adapted prepare a number of other
compounds of the invention, and alternative methods for preparing
the compounds of this invention are deemed to be within the scope
of this invention. For example, the synthesis of the
non-exemplified compounds according to the invention may be
successfully performed by modifications apparent to the skilled in
the art, e.g., by appropriately protecting the interfering groups,
by utilizing other suitable reagents known in the art than those
described, and or making routine modifications of reaction
conditions. Alternatively, other reactions disclosed herein or
known in the art will be recognized as having applicability for
preparing other compounds of the invention.
TABLE-US-00002 General abbreviations and symbols g gram mg
milligram ng nano gram L liter mL milliliter mol mole mmol
millimole min minutes h hour .degree. C. degrees Celsius EtOAc
Ethyl acetate HRMS High resolution mass spectrometry % percent
.mu.M micromolar mM millimolar TLC thin-layer chromatography HPLC
high-performance liquid chromatography GCMS gas chromatography-mass
spectrometry LCMS liquid chromatography-mass spectrometry GCFID gas
chromatography-flame ionisation detector SM starting material eq.
equivalent Pd/C Palladium on charcoal nM nanomolar
TABLE-US-00003 Spectroscopic abbreviations and symbols .sup.1H NMR
proton nuclear magnetic resonance spectrum .delta. (ppm) chemical
shift relative to tetramethylsilane (.delta..sub.TMS = 0) s singlet
d doublet dd doublet of doublet t triplet dt doublet of triplets q
quartet m multiplet br broad Hz Hertz J coupling constant ddd
double doublet of doublet MHz Mega Hertz
General Synthetic Procedures
Procedure A:
##STR00125##
[0389] A mixture of compound A1 (1.0 mmol equiv.) and an amine
(R.sup.5--NH.sub.2) (1.0 mmol equiv.) was combined in mixed
solvents of TFE (20 mL):McCN (20 mL)) and stirred at 25.degree. C.
for 1 hr. At the end, the reaction mixture was concentrated and
purified by triturating with n-pentane to afford compound E1.
[0390] To a stirred solution of E1 (1.0 mmol equiv.) in mixed
solvent [DCM (10 mL): TFE (10 mL)] was added TMSCN (3.4 mmol
equiv.) at 25.degree. C. The reaction mixture was stirred for 3 h
at 25.degree. C. and concentrated. Crude material was triturated
with n-pentane to provide (I-C) as solid. A similar procedure can
be followed to prepare target molecules of formula (I) starting
from optionally substituted amines (R.sup.5--NH.sub.2) and/or
pyridine derivatives, wherein the hydroxyl and the aldehyde
functionalities are adjacent to each other.
##STR00126##
[0391] A mixture of 3-hydroxypyridine-4-carboxaldehyde (3 g, 24.39
mmol) and 4-fluoro-3-chloro phenyl amine (3.55 g, 24.39 mmol) was
combined in mixed solvents of TFE (20 mL):MeCN (20 mL) and stirred
at 25.degree. C., for 1 hr. At the end, the reaction mixture was
concentrated and purified by triturating with n-pentane to afford 6
g of 4-{[3-chloro-4-fluorophenylimino]-methyl}-pyridin-3-ol.
##STR00127##
[0392] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol (6 g, 24
mmol) in mixed solvent [DCM (10 mL): TFE (10 mL)] was added TMSCN
(10.5 mL, 84 mmol) at 25.degree. C. The reaction mixture was
stirred for 3 hr at 25.degree. C. and concentrated. Crude material
was triturated with n-pentane to provide 4.9 g of
N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diamine
as a pale pink solid. A similar procedure can be followed to
prepare target molecules of formula (I) starting from from
optionally substituted amines (R.sup.5--NH.sub.2) and/or pyridine
derivatives, wherein the hydroxyl and the aldehyde functionalities
are adjacent to each other.
Procedure B:
##STR00128##
[0394] A mixture of compound Al (1.0 mmol quiv.) and an amine
(R.sup.5--NH.sub.2)(1.0 mmol equiv.) was combined in mixed solvents
of TFE (20 mL):MeCN (20 mL) and stirred at 25.degree. C., for 2 h.
The reaction was monitored by TLC SiO.sub.2. At the end, reaction
mixture was diluted with DCM (10 mL) and TMSCN (3.4 mmol) was
added, and the resulting reaction mixture was stirred for 12 h and
concentrated and triturated with EtOAc/pentane to provide compound
(I-C) as solid. A similar procedure can be followed to
preparetarget molecules of formula (I) starting from optionally
substituted amines (R.sup.5--NH.sub.2) and/or pyridine derivatives,
wherein the hydroxyl and the aldehyde functionalities are adjacent
to each other.
##STR00129##
[0395] A mixture of 3-hydroxy pyridine 2-carboxaldehyde (500 mg,
4.065 mmol) and 3-chloro-4-fluoroaniline (591 mg, 4.065 mmol) was
combined in TFE (10 mL):MeCN (10 mL) and stirred at 25.degree. C.
for 2 hr. The reaction was monitored by TLC SiO2. At the end,
reaction mixture was diluted with DCM and TMSCN (2.6 mL, 21.78
mmol) was added. The resulting reaction mixture was stirred for 12
hr and concentrated and triturated with EtOAc/pentane to provide
450 mg of
N.sup.3-(3-chloro-4-fluorophenyl)-furo[3,2-b]pyridine-2,3-diamine.
A similar procedure can be followed to prepare target molecules of
formula (1) starting from optionally substituted amine
(R.sup.5-NH.sub.2) and/or pyridine derivatives, wherein the
hydroxyl and the aldehyde functionalities are adjacent to each
other.
Procedure C:
##STR00130##
[0397] A mixture of compound Al (1.0 mmol eq.) and an amine
(R.sup.5--NH.sub.2) (1 mmol eq.) was combined in solvent mixtures
of TFE (20 mL):MeCN (20 mL)) and stirred at 25.degree. C., for 2 h.
The reaction was monitored by TLC. At the end of reaction period,
reaction mixture was diluted with DCM (10 mL). Next, TMSCN (3.4
mmol eq.) followed by TMSOTf (0.2 mmol eq.) were added to the
resulting reaction mixture. The reaction mixture was stirred for 12
h and concentrated under reduced pressure to provide a crude mass
which was purified by trituration with EtOAc/pentane forming
compound (I-C). A similar procedure can be followed to prepare
target molecules of formula (I) starting from from optionally
substituted amines (R.sup.5--NH.sub.2) and/or pyridine derivatives,
wherein the hydroxyl and the aldehyde functionalities are adjacent
to each other.
Procedure D:
##STR00131##
[0399] The compound QQQ1 was prepared according to scheme 16
(European Journal of Medicinal Chemistry, 2009, 44, 1893-1899). To
a stirred solution of bromo-ester compound QQQ1 (1.0 mmol eq.)
dissolved in toluene (5 mL) was added an amine (R.sup.5--NH.sub.2)
(1.4 mmol eq.), Cs.sub.2CO.sub.3 (1.5 mmol eq.), BINAP (0.2 mmol
eq.) and degassed for 20 min. Next, Pd.sub.2(dba).sub.3 (0.048 mmol
eq.) was added to the reaction mixture. The reaction mixture was
heated at 100-105.degree. C. for 16 h. The reaction mixture was
filtered through a bed of Celite and washed with ethyl acetate. The
filtrate was diluted with water and extracted with EtOAc. The
combined organic layer was dried over sodium sulfate and further
concentrated under reduced pressure resulting in the formation of a
crude mass. The resulting mass was purified by column
chromatography on silica gel using solvent mixtures of EtOAc and
hexane as eluent resulting in the formation of the compound JJJ1 as
a solid.
[0400] Next, the compound JJJ1 was dissolved in THF (10 mL), TEA
(2.5 mmol eq.), and DMAP (0.6 mmol eq.) at 0.degree. C.-5.degree.
C. After 10 minutes, Boc-anhydride (2.2 mmol eq.) was added
dropwise to the reaction mixture and heated at 50.degree.
C.-60.degree. C. for 4 h. After completion of the reaction, water
was added to the reaction mixture. The reaction mixture was further
extracted with EtOAc. The combined organic layer was dried over
sodium sulfate, concentrated under reduced pressure resulting in
the formation of a crude mass which was purified by column
chromatography on silica gel using solvent mixtures of EtOAc and
hexanes as eluent to form Boc-protected compound.
[0401] The Boc-protected compound was dissolved in a
solvent-mixture of THF (15 mL):MeOH (9 mL):H.sub.2O (3 mL) to which
LiOH (2.0 mmol eq.) was added at 0.degree. C.-5.degree. C. The
reaction mixture was continuously stirred at 0.degree. C.-5.degree.
C. for 16 h. Next, the reaction mixture was concentrated under
reduced pressure. The resulting residue was diluted with water and
acidified with 5% citric acid solution (pH=4.0). The resulting
solid precipitate was filtered, washed with water and dried under
vacuum forming a corresponding carboxylic acid.
[0402] The resulting carboxilic acid was dissolved in tert-BuOH (10
mL). Next DPPA (1.2 mmol eq.) and DIPEA (1.1 mmol eq.) were added
to the aforementioned solution at room temperature and the
resulting reaction mixture was heated at 100.degree. C. for 2 h.
After completion of reaction, the mixture was concentrated under
reduced pressure to form a residue which was diluted with water,
neutralized with saturated NaHCO.sub.3 solution, and extracted with
EtOAc. The combined organic layer was dried over sodium sulfate and
concentrated under reduced pressure resulting in the formation of a
crude mass. The mass was purified by column chromatography on
silica gel using solvent mixture of MeOH and DCM as eluent to form
the corresponding carbamate.
[0403] The resulting carbamate was dissolved in dioxane to which
dioxane-HCl solution (4 M; 9.0 mmol eq) was added at 0.degree. C.
5.degree. C. The resulting mixture was stirred for 8 h at
25.degree. C. After completion of the reaction, the reaction mass
was concentrated under reduced pressure resulting in the formation
of a residue which was diluted with DCM and basified with liq.
NH.sub.3. The DCM layer was dried over sodium sulfate, concentrated
under reduced pressure forming a crude mass which was subsequently
purified by column chromatography on silica gel using solvent
mixtures of MeOH and DCM as eluent resulting in the formation of a
compound (I-JJ) as a solid.
Procedure E:
##STR00132##
[0405] The compound LLL1 was prepared according to scheme 15. A
solution of compound LLL1 (1.0 mmol eq.), R.sup.2-substituted
aryl/heteroarylboronic acid/ester (1.05 mmol eq.),
Pd(PPh.sub.3).sub.4, (0.05 mmol eq.), potassium carbonate (1.5 mmol
eq.) in toluene:water (1:1 mL) mixture was refluxed overnight at
100.degree. C. After completion of the reaction, the mixture was
cooled to room temperature, diluted with ethyl acetate, filtered
through a Celite bed, and washed with ethyl acetate. The combined
filtrates were washed with water and brine. The organic layer was
dried over sodium sulfate and concentrate under reduced pressure to
give residue which was purified by column chromatography using
hexane as eluent on silica gel to form compound FFF1 as a
solid.
[0406] The compound FFF1 was dissolved in mixture of
chloroform:acetic acid (1:1 mL) and cooled to 0.degree. C.
5.degree. C. To the above solution, NBS (1.26 mmol eq.) was added
portion wise. The reaction mixture was stirred at room temperature
for 48 h. After completion of the reaction, the mass was quenched
by a saturated solution of sodium thiosulfate and extracted with
chloroform. The combined organic layers were washed with saturated
solution of sodium bicarbonate and brine, dried over sodium sulfate
and concentrated under reduced pressure forming a residue. The
residue was purified by column chromatography using hexane as
eluent on silica gel forming a corresponding 3-bromo-compound.
[0407] The resulting 3-bromo-compound was dissolved in acetic
anhydride (12.7 mmol eq.)the solution was cooled to 0.degree. C. A
mixture of fuming nitric acid (6.8 mmol eq.) in acetic acid (3.4
mmol eq.) was added to the above solution and the resulting mass
was stirred for 2 h. After completion of reaction, the mixture was
quenched in ice-cold water, extracted with DCM, dried over
anhydrous sodium sulfate, and concentrated under reduced pressure
and formed a residue. The resulting residue was purified by column
chromatography using hexane as eluent on silica gel to provide a
corresponding 2-bromo-3-nitro compound as solid.
[0408] Next, the 2-bromo-3-nitro compound was dissolved in DMF (10
mL) to which an amine (R.sup.5--NH.sub.2) (2.0 mmol eq.) was added.
The reaction mixture was heated at 120.degree. C. for 1 h. After
completion of the reaction, the mixture was poured into ice-cold
water while stirring resulting in the precipitation of a solid. The
solid precipitate was filtered and washed with water. The
precipitate was next dried under vacuum and recrystallized with a
mixture of DCM and hexane resulting in the formation of compound
III1.
[0409] Activated 10% Pd/C was added under nitogen gas at room
temperature to a solution of compound III1 in methanol. The
reaction mass was stirred for 3-4 h under hydrogen gas (balloon).
The reaction mixture was filtered through a Celite bed under
nitrogen and washed with methanol. The filtrate was distilled
generating a residue which was then crystallized with a DCM and
hexane mixture to form compound (I-HH) as a solid.
[0410] Procedure F:
##STR00133##
[0411] To a stirred solution of compound RRR1 (1.0 mmol eq.) in THF
(2 mL) and methanol (2 mL) was added zinc dust (2.0 mmol eq.) and
NH.sub.4Cl (1.0 mmol eq.) at room temperature. The mixture was
stirred for 3 h. The mixture was filtered through a pad of Celite
and washed with ethyl acetate. The filtrate was diluted with EtOAc,
and then washed with water and brine. The organic layer was dried
over sodium sulfate, concentrated under reduced pressure generating
a crude mass. The mass was purified by triturating with MTBE and
pentane to form a compound (I-KK) as a solid.
Procedure G:
##STR00134##
[0413] To a stirred solution of compound YYY1 (1.0 mmol eq.) in THF
was added pyridine (1.3 mmol eq.) followed by the addition of ethyl
chloroformate (1.1 mmol eq.) at 25.degree. C. The reaction mixture
was stirred for 3-10 h at 25.degree. C. and concentrated. The crude
mass was dissolved in EtOAc and washed first with water followed by
a second wash with brine. The mass was next dried over
Na.sub.2SO.sub.4 and concentrated. The mass was subjected to
Prep-TLC/trituration to form a solid monocarbamate compound ZZZ1A
with a trace of dicarbamate.
Procedure H:
##STR00135##
[0415] To a stirring solution of compound YYY1 (1.0 mmol eq.)
dissolved in THF was added pyridine (10.0 mmol eq.) followed by
ethyl chloroformate (8.0 mmol eq.) at 25.degree. C. Reaction
mixture was stirred for 20 h at 25.degree. C. and concentrated. The
crude material was dissolved in EtOAc, washed thoroughly with water
followed by brine, dried over Na.sub.2SO.sub.4 and concentrated.
The crude mass was subjected to Prep-TLC/trituration to afford the
major desired dicarbamate compound ZZZ1B along with monocarbamate
as a solid.
Procedure I:
##STR00136##
[0417] The compound WWWWW1 was prepared according to scheme 27.
Briefly, to a stirred solution of compound WWWWW1 (1.0 mmol eq.) in
DMF (5 mL) was added an amine (R.sup.5--NH.sub.2) (3.0 mmol eq.)
(0.672 g, 4.62 mmol) at room temperature. The reaction mass was
heated at 100-120.degree. C. and stirring continued for 12 h. After
completion of reaction, the reaction mass was quenched with
ice-water. The solid was precipitated, filtered and washed with
water to generate a wet solid mass which was dried under hot air
oven to afford compound ,0000(1 as a solid.
[0418] The compound XXXXX1 was dissolved in a mixture of DMF (5 mL)
and H.sub.2O (1 mL). R.sup.4-substituted aryl/heteroarylboronic
acid/ester (1.3 mmol eq.), K.sub.3PO.sub.4 (3.0 mmol eq.) and
Pd(PPh.sub.3).sub.4 (0.09 mmol eq.) were added to XXXXX1 at room
temperature. The resulting reaction mixture was stirred at
100-120.degree. C. for 2 h. Completion of the reaction was
monitored by TLC. After completion of reaction, the reaction mass
was cooled to room temperature and filtered through Celite bed. The
filtrate was extracted with ethyl acetate. The extract was washed
with brine and dried over sodium sulfate to remove moisture. The
extract as further concentrated under reduced pressure to remove
solvents yielding a crude mass which was purified by column
chromatography on silica gel using solvent mixture of EtOAc/hexane
as eluent to form YYYYY1 as a solid.
[0419] 10% Pd/C was added under nitrogen gas at room temperature to
a stirring solution of compound YYYYY1 (1.0 mmol eq.) dissolved in
methanol (8 mL). Then nitrogen gas was replaced by hydrogen gas
balloon and stirring was further continued at ambient temperature
for 3-4 h. After completion of reaction, the reaction mixture was
filtered through Hyflo bed and washed with methanol. The filtrate
was evaporated under reduce pressure to give rise to a crude mass
which was purified by trituration using pentane to form a compound
(I-CCC) as a solid.
Procedure J:
##STR00137##
[0421] An amine (R.sup.5--NH.sub.2) (1 mmol eq.), TMSCN (5.2 mmol
eq.), and TMSOTf (0.2 mmol eq.) were added at room temperature in a
sealed tube to a stirred solution of compound GGGGG1 (1 mmol eq.)
in DCM (6.0 mL). The reaction mixture was stirred for 1 h at
40.degree. C., followed by addition of 10 mmol NH.sub.4OAc buffer
(3.0 mL). The reaction mixture was stirred for 12 h and was
filtered through a sintered funnel. The filtered solid was washed
with MTBE, hexanes, Ethyl Acetate or mixtures thereof to remove
trace impurities to generate compound (I-UU) as solid.
Procedure K:
##STR00138##
[0423] The compound KKKK1 was prepared according to scheme 18.
Briefly, to a stirred solution of KKKK1 (1.0 mmol eq.) in dioxane
(1 mL) was added dropwise R.sup.5substituted thiophenol or phenol
in a mixture of solution of NaOH (1.1 mmol eq.), H2O (1 mL) and
dioxane (10 mL) at 0.degree. C., and the reaction mixture was
stirred for 1-2 h. Reaction mass was diluted with water, extracted
with ethyl acetate and washed with brine. The organic layers were
dried over sodium sulfate, concentrated under reduced pressure to
give residue which was purified by column chromatography on silica
gel using suitable mixture of solvents to afford GGGG1 as
solid.
[0424] 10% Pd/C was added under nitrogen gas at room temperature to
a stirring solution of compound GGGG1 (1.0 mmol eq.) dissolved in
ethyl acetate (8 mL). Then, nitrogen gas was replaced by hydrogen
gas balloon and stirring was further continued at ambient
temperature for 10-12 h. After completion of reaction, the reaction
mixture was filtered through Hyflo bed and washed with methanol.
The filtrate was evaporated under reduced pressure to form to a
crude mass which was purified by column chromatography on silica
gel using solvent mixtures to form an amine compound. The amine
compound was dissolved in diethyl ether (5 mL). To this solution
was added a saturated solution (10 mL) of HCl in diethyl ether. The
reaction mass was stirred at room temperature resulting in
precipitation of a solid. The precipitate was separated by
decantation and the solid precipitate was dried under vacuum to
form compound (I-B).
Example 1
Synthesis of N.sup.3-(3-chloro-4-fluorophenyl)furo
pyridine-2,3-diamine (Compound 1)
##STR00139##
[0426] 3-Hydroxypyridine-2-carboxaldehyde (500 mg, 4.065 mmol) was
taken in mixed solvent (TFE (10 mL): MeCN (10 mL)) and
3-chloro-4-fluoroaniline (591 mg, 4.065 mmol) was added to it at
25.degree. C., resulting mixture was stirred at this temperature
for 2 hr. TLC indicated no SM remained, to this imine was added
TMSCN (2.6 mL, 21.138 mmol) at 25.degree. C. After stirring the
reaction mixture for 12 hr at 25.degree. C., it was concentrated
and triturated with EtOAc/pentane to yield 450 mg of
N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[3,2-b]pyridine-2,3-diamine
(39% Yield) LCMS: 278(M+H), HPLC: 98.64%, .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 8.07 (d,1H, J=4.8 Hz), 7.52 (d,
1H, J=7.8 Hz), 7.12-7.07 (m, 2H,), 6.87-6.85 (m, 1H), 6.77 (s, 2H),
6.56-6.55 (m, 1H), 6.51-6.49 (m, 1H).
Example 2
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diamine
(Compound 2)
##STR00140##
[0427] Step 1: 3-Methoxymethoxy-pyridine
##STR00141##
[0429] To a stirred solution of 3-hydroxypyridine (60 g, 662.9
mmol) in THF:DMF (120:280 mL) at 0.degree. C. was added t-BuOK
(81.8 .mu.m, 729.28 mmol) portion-wise. After stirring the reaction
mixture for 15 min, methoxymethyl chloride (52 mL, 696.13 mmol) was
added to it at 0.degree. C. and the resulting mixture was stirred
for 1 hr at 25.degree. C. Reaction mixture was diluted with water
and extracted with ethyl acetate (4.times.500 mL). The organic
layer was dried over anhydrous sodium sulfate, concentrated under
reduced pressure to afford 100 g crude which was purified by column
chromatography using silica (100-200 mesh) and 10% EtOAc-hexane as
eluent to afford 3-methoxymethoxy-pyridine (54 g) as pale brown
liquid. LCMS: 140 (M+H).
Step 2: 3-Methoxymethoxy-pyridine-4-carbaldehyde
##STR00142##
[0431] To a stirred solution of 3-methoxymethoxypyridine (2 g,
14.3885 mmol) in anhydrous THF (40 mL) was added TMEDA (1.83 g,
15.82 mmol) at 25.degree. C. The reaction mixture was cooled to
-78.degree. C., n-BuLi (7.3 mL, 15.82 mmol, 2.17 M in hexane) was
added dropwise manner maintaining the temperature -78.degree. C.
After stirring for 2 hr at -78.degree. C., DMF (1.52g, 20.86 mmol)
was added to it and stirred for 2 hr at 25.degree. C. Reaction
mixture was cooled to -40.degree. C. and saturated ammonium
chloride solution was added drop wise. The reaction mass was
extracted with ethyl acetate (250 mL.times.2), EtOAc part was
washed with water followed by brine, dried over sodium sulfate and
concentrated under reduced pressure to afford 3 g of crude product
which was passed through a pad of silica (100-200 mesh) using 10%
EtOAc-hexane as eluent to afford 1.6 g of
3-methoxymethoxy-pyridine-4-carbaldehyde as pale yellow liquid.
GC-MS: 167 (m/z).
Step 3: 3-Hydroxy-pyridine-4-carbaldehyde
##STR00143##
[0433] To a stirred solution of
3-methoxymethoxypyridine-4-carbaldehyde (11 g, 65.83 mmol) in THF
(50 mL) was added 3N HCl (100 mL) and stirred at 60.degree. C. for
1 hr. The reaction mixture was cooled under ice bath and pH was
adjusted to 7 with solid K.sub.2CO.sub.3. Resulting mixture was
extracted with EtOAc (250 mL.times.5). The organic layer was dried
over sodium sulfate, concentrated under reduced pressure to afford
15g of crude which was purified by column chromatography using
silica gel (100-200 mesh) and 23% EtOAc/hexane as eluent to afford
4 g of 3-hydroxy-pyridine-4-carbaldehyde as pale yellow solid.
GC-MS: 123 (m/z), .sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta.
11.04 (bs, 1H), 10.37 (s, 1H), 8.46 (s, 1H), 8.20 (d, 1H, J=4.88
Hz), 7.46 (d, 1H, J=4.88Hz). GC-FID: 99.51%.
Step 4: 4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol
##STR00144##
[0435] 3-Hydroxypyridine-4-carbaldehyde (3 g, 24.39 mmol) was taken
in mixed solvent (TFE (20 mL):MeCN(20 mL)) and
4-fluoro-3-chloroaniline (3.55 g, 24.39 mmol) was added to it at
25.degree. C. The resulting mixture was stirred at this temperature
for 1 hr. The reaction mass was concentrated and purified by
triturating with n-pentane to afford 6 g of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-o 1) LCMS:
251.2 (M+H).
Step 5:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diamine
##STR00145##
[0437] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol (6 g, 24
mmol) in mixed solvent [DCM (10 mL):TFE (10 mL)] was added TMSCN
(10.5 mL, 84 mmol) at 25.degree. C. The reaction mixture was
stirred 3 hr at 25.degree. C., concentrated, and the crude material
was triturated with n-pentane to provide 4.9 g (73% yield) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diamine
as pale pink solid. LCMS: 278 (M+H), HPLC: 98.65%, .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 8.41 (s, 1H), 8.06 (d, 1H,
J=5.08Hz), 7.14-7.10 (m, 2H), 6.91 (s, 2H), 6.86 (d, 1H, J=5.08
Hz), 6.56-6.54 (m, 1H), 6.48-6.45 (m, 1H).
Example 3
Synthesis of
(2-amino-3-((3-chloro-4-fluorophenyl)amino)-7-methylfuro[2,3-c]pyridin-4--
yl)methanol (Compound 56)
##STR00146##
[0438] Step 1:
4-{[(E)-3-Chloro-4-fluoro-phenylimino]-methyl}-5-hydroxymethyl-2-methyl-p-
yridin-3-ol
##STR00147##
[0440] 3-Hydroxy-5-hydroxymethyl-2-methyl-pyridine-4-carbaldehyde
hydrochloride (250 mg, 1.2277 mmol) was taken in mixed solvent (TFE
(3 mL):MeCN (3 mL)), 4-fluoro-3-chlorophenylamine (178 mg, 1.2277
mmol) was added to it at 25.degree. C., and the resulting mixture
was stirred at this temperature for 3 hr. The reaction mass was
concentrated, dissolved in EtOAc, washed with sodium bicarbonate
solution, dried over Na.sub.2SO.sub.4, and concentrated to afford
100 mg of
4-{[(E)-3-chloro-4-fluoro-phenylimino]-methyl}-5-hydroxymethyl-2-methyl-p-
yridin-3-ol. LCMS: 295 (M+H)
Step 2:
(2-amino-3-((3-chloro-4-fluorophenyl)amino)-7-methylfuro[2,3-c]pyr-
idin-4-yl)methanol
##STR00148##
[0442] To a stirred solution of
4-{[(E)-3-chloro-4-fluoro-phenylimino]-methyl}-5-hydroxymethyl-2-methyl-p-
yridin-3-ol (100 mg, 0.3401 mmol) in mixed solvent [DCM (2 mL): TFE
(2 mL)] was added TMSCN (0.149 mL, 1.19 mmol) at 25.degree. C. The
reaction mixture was stirred for 4 hr at 2.degree. C. and
concentrated. The crude material was triturated with n-pentane
followed by MTBE to yield 30 mg (27% yield) of
[2-amino-3-(3-chloro-4-fluoro-phenylamino)-7-methyl-furo[2,3-c]pyridin-4--
yl]-methanol. LCMS: 322 (M+H), HPLC: 98.82%, .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 7.94 (s, 1H), 7.12 (t, 1H, J=9
Hz), 6.89 (s, 1H), 6.78 (s, 2H), 6.54-6.53 (m, 1H), 6.45-6.43 (m,
1H), 4.92 (s, 1H), 4.45 (s, 2H), 2.46 (s, 3H).
Example 4
Synthesis of
4-Chloro-N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diami-
ne (Compound 57)
##STR00149##
[0443] Step 1: 3-chloro-5-methoxymethoxy-pyridine
##STR00150##
[0445] To a stirred solution of 3-chloro-5-hydroxypyridine (500 mg,
3.86 mmol) in THF:DMF (1 mL:2.3 mL) at 0.degree. C. was added SuOK
(480 mg, 4.24 mmol) portion-wise. After stirring the reaction
mixture for 15 min, methoxymethyl chloride (320 mg, 4.05 mmol) was
added to it at 0.degree. C. and the resulting mixture was stirred
for 0.5 hr at 25.degree. C. The reaction mixture was diluted with
water and extracted with ethyl acetate (2.times.50 mL). The organic
layer was dried over anhydrous sodium sulfate, concentrated under
reduced pressure to afford the crude which was purified by column
chromatography using silica (100-200 mesh) and 10% EtOAc-hexane as
eluent to afford 3-chloro-5-methoxymethoxy-pyridine (200 mg) as
pale brown liquid.
Step 2: 3-Chloro-5-methoxymethoxy-pyridine-4-carbaldehyde
##STR00151##
[0447] To a stirred solution of 3-chloro-5-methoxymethoxypyridine
(500 mg, 2.89 mmol) in anhydrous THF (5 mL) was added LDA (1 M
soln, 4.05 mL) at -78.degree. C. After stirring for 30 min at
-78.degree. C., N-formyl-piperidine (650 mg, 5.78 mmol) was added
to it and stirred for 2 hr at -78.degree. C. The reaction mixture
was quenched with water, extracted with ethyl acetate (50
mL.times.2). The EtOAc part was washed with water followed by
brine, dried over sodium sulfate and concentrated under reduced
pressure to afford the crude which was passed through a pad of
silica (100-200 mesh) using 10% EtOAc-hexane as eluent to afford
250 mg of crude 3-chloro-5-methoxymethoxy-pyridine-4-carbaldehyde
as pale yellow liquid. GCMS: 201 (m/z).
Step 3: 3-Chloro-5-hydroxy-pyridine-4-carbaldehyde
##STR00152##
[0449] To a stirred solution of
3-chloro-5-methoxymethoxypyridine-4-carbaldehyde (450 mg, 2.2388
mmol) in THF (4 mL) was added 3N HCl (4 mL) and stirred at
60.degree. C. for 2 hr. The reaction mixture was cooled under ice
bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (50 mL.times.3). The
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford the crude which was purified by column
chromatography using silica gel (100-200 mesh) and 7% EtOAc/hexane
as eluent to afford 110 mg of
3-chloro-5-hydroxy-pyridine-4-carbaldehyde. LCMS: 156 (M-H).
Step 4:
5-Chloro-4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol
##STR00153##
[0451] 3-Chloro-5-hydroxy-pyridine-4-carbaldehyde (110 mg, 0.69
mmol) was taken in mixed solvent [TFE (1.5 mL):MeCN(1.5 mL)] and
4-fluoro-3-chloro phenyl amine (100 mg, 0.69 mmol) was added to it
at 25.degree. C., and the resulting mixture was stirred at this
temperature for 2 hr. The reaction mass was concentrated and
purified by triturating with n-pentane to afford 100 mg of
5-chloro-4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol.
LCMS: 285 (M+H).
Step 5:
4-Chloro-N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,-
3-diamine
##STR00154##
[0453] To a stirred solution of
5-chloro-4-{[3-chloro-4-fluoro-phenylimino]-methyl}-pyridin-3-ol
(100 mg, 0.35 mmol) in mixed solvent [DCM (1.5 mL): TFE (1.5 mL)]
was added TMSCN (120 mg, 1.23 mmol) at 25.degree. C. The reaction
mixture was stirred for 3 hr at 25.degree. C., concentrated, and
crude material was triturated with DCM/MeCN to yield 20 mg (18%
yield) of
4-chloro-N.sup.3-(3-chloro-4-fluoro-phenyl)-furo[2,3-c]pyridine-2,3-diami-
ne as off white solid. LCMS: 312 (M+H), HPLC: 97.01%, .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 8.37 (s, 1H), 8.03 (s, 1H), 7.25
(s, 2H), 7.11(t, 1H, J=9.04 Hz), 7.06 (s, 1H), 6.58-6.57 (m, 1H),
6.47-6.45 (m, 1H).
Example 5
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-5-methoxy-furo[2,3-c]pyridine-2,3-diam-
ine (Compound 64)
##STR00155##
[0454] Step 1: 5-Bromo-2-methoxy-pyridine
##STR00156##
[0456] To a stirred solution of 2-methoxypyridine (2 g, 18.33 mmol)
in MeCN (54 mL) was added NBS (3.9 g, 21.998 mmol) at 0.degree. C.
The reaction mixture was stirred for 16 hr. The reaction mass was
filtered through a pad of silica and the filtrate was evaporated to
provide the crude product. Column chromatography rendered 2 g of
5-bromo-2-methoxypyridine.
Step 2:
2-Methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-
e
##STR00157##
[0458] 5-Bromo-2-methoxypyridine (5 g, 26.59 mmol),
bis(pinacolato)diborane (10.13 g, 39.89 mmol) and potassium acetate
(10.44 g, 106 mmol) were taken in dry toluene (60 mL) and degassed
with nitrogen for 20 min. Pd(dppf)Cl.sub.2. DCM (2.17 g, 2.66 mmol)
was added to the reaction under nitrogen atmosphere and the
resulting mixture was refluxed for 2 hr. The reaction progress was
monitored by TLC. After completion of the reaction, the mixture was
cooled to 25.degree. C. and filtered through a Celite.RTM. reagent
pad. Filtrate was diluted with ethyl acetate (200 mL), washed with
water followed by brine, dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude which was
purified by silica gel (100-200 mesh) column chromatography using
10% EtOAc in hexane as eluent to afford 3.6 g of
2-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine.
Step 3: 6-Methoxy-pyridin-3-ol
##STR00158##
[0460] To a stirred suspension of sodium perborate tetrahydrate
(6.87 g, 44.68 mmol) in water was added
2-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
(3.5 g, 14.68 mmol) in THF (70 mL) at room temperature. The
resulting reaction mixture was stirred for 2 hr at room
temperature. The reaction mass was extracted with ethyl acetate
(200 mL), washed with brine dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude which was
purified by silica gel (100-200 mesh) column chromatography using
20% EtOAc in hexane as eluent to afford 2.4 g of
6-methoxy-pyridin-3-ol. LCMS: 126 (M+H).
Step 4: 2-Methoxy-5-methoxymethoxy-pyridine
##STR00159##
[0462] To stirred solution of 6-methoxypyridin-3-ol (2.4 g, 19.20
mmol) in THF:DMF (4 mL: 9 mL) was added potassium tert-butoxide
(2.37 g, 21.12 mmol) at 0.degree. C. The reaction mixture was
stirred for 15 min at 0.degree. C. and methoxymethylene chloride
(1.6 g, 20.16 mmol) was added dropwise at 0.degree. C. The reaction
mixture was allowed to stir 40 min at room temperature. The
reaction mass was diluted with ethyl acetate (100 mL), washed with
water followed by brine dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude which was
purified by silica gel (100-200 mesh) column chromatography by
using 10% EtOAc in hexane as eluent to afford 2 g of
2-methoxy-5-methoxymethoxypyridine. LCMS: 170 (M+H).
Step 5: 2-Methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde
##STR00160##
[0464] To a stirred solution of 2-methoxy-5-methoxymethoxypyridine
(1.7g, 10.05 mmol) in THF (17 mL) was added TMEDA (1.65 mL, 11.05
mmol) and reaction mixture was cooled to -78.degree. C. n-BuLi
(2.17 M, 5.09 mL) was then added in dropwise at -78.degree. C. Then
reaction mixture was stirred for 1 hr at -78.degree. C. DMF (1.2
mL, 14.57 mmol) was then added in dropwise at -78.degree. C. Then
reaction mixture was stirred for 1 hr at -78.degree. C. The
reaction progress was monitored by TLC. The reaction mixture was
cooled to room temperature and extracted with ethyl acetate (100
mL). The combined organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to crude material. The crude
material was purified by silica gel (100-200 mesh) column
chromatography by eluting with 8% EtOAc in hexane. The collected
fractions were evaporated to afford 1.4 g of
2-methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde. GCMS: 197
(m/z).
Step 6: 5-Hydroxy-2-methoxy-pyridine-4-carbaldehyde
##STR00161##
[0466] To a stirred solution of
2-methoxy-5-methoxymethoxypyridine-4-carbaldehyde (1.5 g, 7.61
mmol) in THF (3 mL) was added 3N HCl (15 mL) at room temperature
and the reaction mixture was heated to 70.degree. C. for 2 hr. The
reaction mixture was cooled under ice bath, neutralized with
K.sub.2CO.sub.3 and extracted with ethyl acetate (50 mL.times.2).
The combined organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude product
which was purified by column chromatography to afford 600 mg of
5-hydroxy-2-methoxy-pyridine-4-carbaldehyde.
[0467] .sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.33 (s, 1H),
10.30 (s, 1H), 8.02 (s, 1H), 6.88 (s, 1H), 3.79 (s, 3H).
Step 7:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-6-methoxy-pyridin-3-ol
##STR00162##
[0469] 5-Hydroxy-2-methoxypyridine-4-carbaldehyde (250 mg, 1.63
mmol) was taken in mixed solvent (TFE (2.5 mL): MeCN (2.5 mL)) and
4-fluoro-3-chlorophenyl amine (240 mg, 1.63 mmol) was added to it
at 25.degree. C. and the resulting mixture was stirred at this
temperature for 2 hr. The reaction mass was concentrated and
purified by triturating with n-pentane to afford 350 mg of
4-{-chloro-4-fluoro-phenylimino-methyl}-6-methoxy-pyridin-3-ol.
LCMS: 281 (M+H).
Step 8:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-5-methoxy-furo[2,3-c]pyridine-2-
,3-diamine
##STR00163##
[0471] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-6-methoxy-pyridin-3-ol
(350 mg, 1.25 mmol) in mixed solvent [DCM (3.5 mL): TFE (3.5 mL)]
was added TMSCN (0.6 mL, 4.37 mmol) at 25.degree. C. The reaction
mixture was stirred 8 hr at 25.degree. C., concentrated, and crude
material was triturated with n-pentane to provide 70 mg (18% Yield)
of
N.sup.3-(3-chloro-4-fluoro-phenyl)-5-methoxy-furo[2,3-c]pyridine-2,3-diam-
ine as off white solid. LCMS: 308 (M+H), HPLC: 99.02%, .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 7.97 (s, 1H), 7.12 (t, 1H, J=9.1
Hz), 7.02 (s, 1H), 6.93 (s, 2H), 6.55-6.53 (m, 1H), 6.47-6.44 (m,
1H), 6.06 (s, 1H), 3.75 (s, 3H).
Example 6
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-methyl-furo[2,3-c]pyridine-2,3-diami-
ne (Compound 66)
##STR00164##
[0472] Step 1: 3-Methoxymethoxy-2-methyl-pyridine
##STR00165##
[0474] To a stirred solution of 3-hydroxy-2-methyl pyridine (1.5 g,
13.745 mmol) in DCM (20 mL) at 0.degree. C. was added DIPEA (2.8
mL, 16.494 mmol). After stirring the reaction mixture for 10 min,
methoxymethyl chloride (1.2 mL, 16.494 mmol) was added to it at
0.degree. C. and the resulting mixture was stirred for 16 hr at
25.degree. C. The reaction mixture was diluted with water and
extracted with DCM (2.times.50 mL). The organic layer was dried
over anhydrous sodium sulfate and concentrated under reduced
pressure to afford crude material which was purified by column
chromatography using silica (100-200 mesh) and 10% EtOAc-hexane as
eluent to afford 3-methoxymethoxy-2-methyl-pyridine (1.1 g) as pale
yellow liquid. LCMS: 154 (M+H).
Step 2: 3-Methoxymethoxy-2-methyl-pyridine-4-carbaldehyde
##STR00166##
[0476] To a stirred solution of 3-methoxymethoxy-2-methyl-pyridine
(1.1 g, 7.1895 mmol) in anhydrous THF (20 mL) was added TMEDA (1.1
mL, 7.9084 mmol) at 25.degree. C. The reaction mixture was cooled
to -78.degree. C., n-BuLi (3.6 mL, 7.9084 mmol, 2.17 M in hexane)
was added dropwise maintaining the temperature -78.degree. C. After
stirring for 2 hr at -78.degree. C., DMF (0.80 mL, 10.4247 mmol)
was added to it and stirred for 1 hr at -78.degree. C. The
temperature was then slowly raised to 25.degree. C. The reaction
mixture was quenched with saturated ammonium chloride solution,
extracted with ethyl acetate (30 mL.times.2), EtOAc part was washed
with water followed by brine, dried over sodium sulfate and
concentrated under reduced pressure to afford crude material which
was passed through a pad of silica (100-200 mesh) using 10%
EtOAc-hexane as eluent to afford 250 mg of crude
3-methoxymethoxy-2-methylpyridine-4-carbaldehyde as pale yellow
liquid. GCMS: 181 (m/z).
Step 3: 3-Hydroxy-2-methylpyridine-4-carbaldehyde
##STR00167##
[0478] To a stirred solution of
3-methoxymethoxy-2-methylpyridine-4-carbaldehyde (250 mg, 1.3812
mmol) in THF (0.5 mL) was added 3N HCl (2.5 mL) and stirred at
60.degree. C. for 2 hr. The reaction mixture was cooled under ice
bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (20 mL.times.2). The
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford crude which was purified by column
chromatography using silica gel (100-200 mesh) and 23% EtOAc/hexane
as eluent to afford 100 mg of
3-hydroxy-2-methyl-pyridine-4-carbaldehyde as pale yellow solid.
GCMS: 137 (m/z).
Step 4:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-2-methyl-pyridin-3-ol
##STR00168##
[0480] 3-Hydroxy-2-methylpyridine-4-carbaldehyde (100 mg, 0.7299
mmol) was taken in mixed solvent [TFE (1 mL):MeCN (1 mL)] and
4-fluoro-3-chlorophenyl amine (106 mg, 0.7299 mmol) was added to it
at 25.degree. C. The resulting mixture was stirred at this
temperature for 3 hr. The reaction mass was concentrated and
purified by triturating with n-pentane to afford 150 mg of
4-{[3-chloro-4-fluoro-phenylimino ]-methyl}-2-methyl-pyridin-3-ol.
LCMS: 262.8 (M-H).
Step 5:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-methyl-furo[2,3-c]pyridine-2,-
3-diamine
##STR00169##
[0482] To a stirred solution of 4-{[3-chloro-4-fluoro
-phenylimino]-methyl}-2-methyl-pyridin-3-ol (150 mg, 0.5681 mmol)
in mixed solvent [DCM (2 mL): TFE (2 mL)] was added TMSCN (0.25 mL,
1.989 mmol) at 25.degree. C. Reaction mixture was stirred for 5 hr
at 25.degree. C., concentrated, and the crude material was
triturated with MeCN/pentane to yield 40 mg (24% Yield) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-methyl-furo[2,3-c]pyridine-2,3-diami-
ne as reddish brown solid. LCMS: 292 (M+H), HPLC: 99.06%,
.sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta. 7.93 (d, 1H, J=5.1
Hz), 7.13-7.08 (m, 2H), 6.82 (s, 2H), 6.71 (d, 1H, J=5.1 Hz),
6.54-6.52 (m, 1H), 6.48-6.45 (m, 1H), 2.49 (s, 3H).
Example 7
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-4-methoxy-furo[2,3-c]pyridine-2,3-diam-
ine (Compound 67)
##STR00170##
[0483] Step 1:
3-Methoxy-5-(4,4,5,5-tetramethyl41,3,21dioxaborolan-2-yl)-pyridine
##STR00171##
[0485] 3-Bromo-5-methoxypyridine (2g, 10.63 mmol),
bis(pinacolato)diborane (4.05 g, 15.98 mmol) and potassium acetate
(4.78 g, 42.55 mmol) were taken in dry toluene (25 mL) and degassed
with nitrogen for 20 min. Pd(dppf)Cl.sub.2. DCM (0.87 g, 0.10 mmol)
was added to the reaction under nitrogen atmosphere and the
resulting mixture was refluxed for 2 hr. The reaction progress was
monitored by TLC. After completion of the reaction, the reaction
mixture was cooled to 25.degree. C. and filtered through a
Celite.RTM. reagent pad. The filtrate was diluted with ethyl
acetate (100 mL) washed with water followed by brine, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to provide
the crude which was purified by silica gel (100-200 mesh) column
chromatography using 10% EtOAc in hexane as eluent to afford 2.6 g
of
3-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
LCMS: 236 (M+H).
Step 2: 5-Methoxy-pyridin-3-ol
##STR00172##
[0487] To a stirred suspension of sodium-perborate-tetrahydrate
(7.86 g, 51.06 mmol) in water was added
3-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
(4 g, 17.02 mmol) in THF/H.sub.2O (80 mL: 80 mL) at room
temperature. The resulting reaction mixture was stirred for 2 hr at
room temperature. The reaction mass was extracted with ethyl
acetate (200 mL), washed with brine dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude product
which was purified by silica gel (100-200 mesh) column
chromatography using 20% EtOAc in hexane as eluent to afford 1.5 g
of 5-methoxypyridin-3-ol. LCMS: 126 (M+H).
Step 3: 3-Methoxy-5-methoxymethoxy-pyridine
##STR00173##
[0489] To stirred solution of 5-methoxypyridin-3-ol (1.9 g, 15.20
mmol) in THF:DMF (4 mL: 9 mL) was added potassium tert-butoxide
(1.86 g, 16.72 mmol) at 0.degree. C. The reaction mixture was
stirred for 15 min at 0.degree. C. and methoxymethylene chloride
(1.28 g, 15.96 mmol) was added dropwise. The reaction mixture was
allowed to stir 40 min at room temperature. The reaction mass was
diluted with ethyl acetate (100 mL) washed with water followed by
brine dried over Na.sub.2SO.sub.4 and concentrated under reduced
pressure to provide the crude product which was purified by silica
gel (100-200 mesh) column chromatography by using 10% EtOAc in
hexane as eluent to afford 1.5 g of
3-methoxy-5-methoxymethoxypyridine. LCMS: 170 (M+H).
Step 4: 3-Methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde
##STR00174##
[0491] To a stirred solution of 3-methoxy-5-methoxymethoxy-pyridine
(1.5 g, 8.86 mmol) in THF (15 mL) was added TMEDA (1.45 mL, 9.75
mmol) and reaction mixture was cooled to -78.degree. C. n-BuLi
(2.17 M, 4.49 mL) was then added dropwise at -78.degree. C. and the
reaction mixture was allowed to stir 1 hr at this temperature. DMF
(1 mL, 12.85 mmol) was then added at -78.degree. C. and the
resulting mixture was stirred for 1 hr at the same temperature. The
reaction mass was quenched with saturated NH.sub.4Cl solution,
extracted with ethyl acetate (100 mL.times.2). The combined organic
layer was dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure to provide the crude product which was purified by
silica gel (100-200 mesh) column chromatography eluting with 8%
EtOAc in hexane to afford 0.7 g of
3-methoxy-5-methoxymethoxy-pyridine-4-carbaldehyde. GCMS: 197
(m/z).
Step 5: 3-Hydroxy-5-methoxy-pyridine-4-carbaldehyde
##STR00175##
[0493] To a stirred solution of
3-methoxy-5-methoxymethoxypyridine-4-carbaldehyde (0.7 g, 3.55
mmol) in THF (3 mL) was added 3N HCl (7 mL) at room temperature and
the reaction mixture was heated to 70.degree. C. for 2 hr. The
reaction mixture was cooled under ice bath, neutralized with
K.sub.2CO.sub.3 and extracted with ethyl acetate (50 mL.times.2).
The combined organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to provide the crude product
which was purified by column chromatography to afford 300 mg of
3-hydroxy-5-methoxypyridine-4-carbaldehyde. GCMS: 153 (m/z).
Step 6:
4-{[-3-Chloro-4-fluoro-phenylimino]-methyl}-4-methoxy-pyridin-3-ol
##STR00176##
[0495] 3-Hydroxy-5-methoxypyridine-4-carbaldehyde (100 mg, 0.65
mmol) was taken in mixed solvent [TFE (1.0 mL):MeCN (1.0 mL)] and
4-fluoro-3-chloro phenyl amine (95 mg, 0.65 mmol) was added at
25.degree. C. The resulting mixture was stirred at this temperature
for 2 hr. The reaction mass was concentrated and purified by
triturating with n-pentane to afford 150 mg of
4-{[-3-Chloro-4-fluoro-phenylimino]-methyl}-4-methoxy-pyridin-3-ol.
LCMS: 281 (M+H).
Step 6:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-4-methoxy-furo[2,3-c]pyridine-2-
,3-diamine
##STR00177##
[0497] To a stirred solution
4-{[-3-chloro-4-fluoro-phenylimino]-methyl}-4-methoxy-pyridin-3-ol
(150 mg, 0.53 mmol) in mixed solvent [DCM (1.5 mL): TFE (1.5 mL)]
was added TMSCN (0.25 mL, 1.87 mmol) at 25.degree. C. The reaction
mixture was stirred 3 hr at 25.degree. C., concentrated, and the
crude material was triturated with DCM/MeCN to yield 60 mg (37%
Yield) of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-4-methoxy-furo[2,3-c]pyridine-2,3-diam-
ine as pale yellow solid. LCMS: 308 (M+H), HPLC: 98.47%,
.sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta. 8.18 (s, 1H), 7.86 (s,
1H), 7.09 (t, 1H, J=9.1 Hz), 6.99 (s, 1H), 6.68 (s, 2H), 6.55-6.52
(m, 1H), 6.47-6.44 (m, 1H), 3.68 (s, 3H).
Example 8
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-4,7-dimethylfuro[2,3-c]pyridine-2,3-dia-
mine (Compound 72)
##STR00178##
[0498] Step 1:
2,2-Dimethyl-4H-[1,3]dioxino[4,5-c]pyridin-5-yl)methanol
hydrochloride
##STR00179##
[0500] Dry HCl was bubbled to a cooled suspension of pyridoxine
hydrochloride (4.0 g) in dry acetone (100 mL) for 1.5 hr. The
solution was stirred for another 1 hr and then kept at cold
condition for overnight. White crystals appeared at this stage
which was separated out and re-crystallization in EtOH to afford
2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridin-5-yl)methanol
hydrochloride (3.70 g, yield 90.9%.) as white solid. HRMS [M+H]:
210.113; .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 7.90 (s, 1H),
5.13 (t, J=5.4 Hz, 1H), 4.86 (s, 2H), 4.40 (d, J=5.4 Hz, 2H), 2.26
(s, 3H), 1.47 (s, 6H).
Step 2:
5-(Chloromethyl)-2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridine
hydrochloride
##STR00180##
[0502] Thionyl chloride (9 mL) was added to a stirred suspension
(3.7 g, 17.683 mmol) of
2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridin-5-yl)methanol
hydrochloride in anhydrous ether (250 mL) at 0.degree. C. After
refluxing for 5 hr, the precipitate was filtered, washed with
ether, and dried under vacuum. The crude product was recrystallized
with boiling absolute ethanol to give
5-(chloromethyl)-2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridine
hydrochloride (2.5 g, yield 80%) as white solid. HRMS [M+H]
228.079; .sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 7.97 (s, 1H),
4.92 (s, 2H), 4.45 (s, 2H), 2.39 (s, 3H), 1.54 (s, 6H).
Step-3: 4-(Hydroxymethyl)-2,5-dimethylpyridin-3-ol
##STR00181##
[0504] A solution of
5-(chloromethyl)-2,2-dimethyl-4H-[1,3]dioxino[4,5-c]pyridine
hydrochloride (1.0 g, 4.403 mmol) in MeOH (10 mL) was hydrogenated
in the presence of Pd/C (100 mg) and anhydrous NaOAc (366 mg, 4.403
mmol) at room temperature under hydrogen for 2 hr. After completion
of reaction, the mixture was filtered off through a Celite.RTM.
reagent bed and the bed was washed with methanol. The filtrate was
collected and concentrated under reduce pressure to give the crude
mass which was diluted with 1N HCl (20 mL) and held overnight at
room temperature. After filtering out a slight precipitate, the
solution was heated at 80.degree. C. for 15 min and concentrated
under reduce pressure up to dryness. The residue was triturated
with ethanol and the product was crystallized by adding diethyl
ether in the ethanol extract. The solid was filtered off and
basified by saturated solution of sodium bicarbonate. The basic
aqueous mass was extracted with ethyl acetate, washed with brine,
dried over anhydrous sodium sulfate and concentrated under reduce
pressure to give crude mass which was purified by column
chromatography using ethyl acetate on silica gel to afford
4-(hydroxymethyl)-2,5-dimethylpyridin-3-ol (470 mg, yield 69.77%)
as white solid. HRMS [M+H] 154.086; .sup.1H-NMR (500 MHz,
DMSO-d.sub.6): .delta. 9.10 (s, 1H), 7.68 (s, 1H), 5.65 (s, 2H),
2.26 (s, 3H), 2.12 (s, 3H).
Step-4: 3-Hydroxy-2,5-dimethylisonicotinaldehyde
##STR00182##
[0506] To a stirred solution of
4-(hydroxymethyl)-2,5-dimethylpyridin-3-ol (470 mg, 3.070 mmol) in
chloroform (20 mL) was added MnO.sub.2 (5.338 g, 61.406 mmol) at
room temperature under nitrogen. The reaction mixture was stirred
at room temperature for 20 hr under nitrogen. The catalyst was
filtered off through a Celite.RTM. reagent bed and washed with
chloroform. The filtrate was concentrated in vacuum. The crude
product was purified by column chromatography on silica gel using
50% ethyl acetate and hexane mixture to afford
3-hydroxy-2,5-dimethylisonicotinaldehyde (200 mg, 44.1%) as white
solid and taken directly to the next step.
Step-5:
N.sup.3-(3-Chloro-4-fluorophenyl)-4,7-dimethylfuro[2,3-c]pyridine--
2,3-diamine
##STR00183##
[0508] To stirred solution of
3-hydroxy-2,5-dimethylisonicotinaldehyde (200 mg, 1.323 mmol) in
MeOH (2 mL) was added 3-chloro-4-fluoroaniline (193 mg, 1.323 mmol)
and two drops of acetic acid at room temperature under nitrogen.
The resulting reaction mixture was stirred at 50.degree. C. for 15
min. Progress of the reaction was monitor by TLC. After consumption
of aldehyde, TMSCN (263 mg, 2.646 mmol) was added dropwise at
50.degree. C. and the reaction mass was stirred for further 20 min.
The reaction mass was cooled to room temperature and solvent was
removed by vacuum distillation. The crude mass was diluted with
ethyl acetate, filtered through a cotton plug and concentrated
under reduced pressure to give the crude product which was
recrystallized by ethyl acetate and hexane mixture to afford
N.sup.3-(3-chloro-4-fluorophenyl)-4,7-dimethylfuro[2,3-c]pyridine-2,3-dia-
mine (80 mg, yield 25%) as off white solid. HRMS [M+H] 306.080;
.sup.1H-NMR (500 MHz, DMSO-d.sub.6): .delta. 7.67 (s, 1H), 7.08 (t,
J=9.0 Hz, 1H), 7.03 (s, 1H), 6.73 (bs, 2H), 6.50-6.48 (m, 1H),
6.42-6.40 (m, 1H), 2.41 (s, 3H), 2.06 (s, 3H).
Example 9
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-ethyl-furo[2,3-c]pyridine-2,3-diamin-
e (Compound 73)
##STR00184##
[0509] Step 1: 2-Chloro-3-methoxymethoxy-pyridine
##STR00185##
[0511] To a stirred solution of 2-chloro-3-hydroxy-pyridine (5 g,
38.59 mmol) in THF: DMF (10:25 mL) at 0.degree. C. was added t-BuOK
(4.763 g, 42.45 mmol) portionwise. After stirring the reaction
mixture for 15 min, methoxymethylchloride (3.062 mL, 40.5 mmol) was
added to it at 0.degree. C. and the resulting mixture was stirred
for 1 hr at 25.degree. C. The reaction mixture was diluted with
water and extracted with ethyl acetate (3.times.150 mL). The
combined organic layer was dried over anhydrous sodium sulfate,
concentrated under reduced pressure to afford crude which was
purified by column chromatography using silica (100-200 mesh) and
10% EtOAc-hexane as eluent to afford
2-chloro-3-methoxymethoxypyridine (5.4 g) as pale brown liquid.
LCMS: 174 (M+H).
Step 2: 2-Ethyl-3-methoxymethoxy-pyridine
##STR00186##
[0513] To a stirred solution of 2-chloro-3-methoxymethoxy-pyridine
(3 g, 17.341 mmol) in DMF (10 mL) was added triethylborate (1.78 g,
18.208 mmol) and K.sub.2CO.sub.3 (3.58 g, 26.011 mmol), the
reaction mass was degassed with argon, Pd(PPh.sub.3).sub.4 (0.5 g,
0.4336 mmol) was then added and heated at 80.degree. C. for 3 hr.
The reaction mass was filtered through Celite.RTM. reagent, the
filtrate was diluted with water and acidified to pH 4 using 1N HCl,
and the resulting mixture was stirred for 15 min. To this
suspension was added saturated NaHCO.sub.3 solution to make a pH 9.
The solution was extracted with MTBE (200 mL.times.2). The combined
organic part was dried over Na.sub.2SO.sub.4, evaporated to dryness
and the crude product was purified by column chromatography to
provide 1.3 g of 2-ethyl-3-methoxymethoxy-pyridine. LCMS: 168
(M+H).
Step 3: 2-Ethyl-3-methoxymethoxy-pyridine-4-carbaldehyde
##STR00187##
[0515] To a stirred solution of 2-ethyl-3-methoxymethoxypyridine (1
g, 5.988 mmol) in anhydrous THF (10 mL) was added TMEDA (1 mL, 6.58
mmol) at 25.degree. C. The reaction mixture was cooled to
-78.degree. C., n-BuLi (2.17 M in hexane; 3.03 mL, 6.58 mmol) was
added dropwise, maintaining the temperature at -78.degree. C. After
stirring for 2 hr at -78.degree. C., DMF (0.67 mL, 8.68 mmol) was
added, the solution stirred for 1 hr at -78.degree. C. and the
temperature slowly raised to 25.degree. C. The reaction mixture was
quenched with saturated ammonium chloride solution, extracted with
ethyl acetate (100 mL.times.2), the EtOAc part was washed with
water followed by brine, dried over sodium sulfate and concentrated
under reduced pressure to afford crude material which was passed
through a pad of silica (100-200 mesh) using 10% EtOAc-hexane as
eluent to afford 900 mg of crude
2-ethyl-3-methoxymethoxy-pyridine-4-carbaldehyde. GCMS: 195
(m/z).
Step 4: 2-Ethyl-3-hydroxy-pyridine-4-carbaldehyde
##STR00188##
[0517] To a stirred solution of
2-ethyl-3-methoxymethoxypyridine-4-carbaldehyde (900 mg, 4.615
mmol) in THF (3 mL) was added 3N HCl (5 mL) and stirred at
60.degree. C. for 2hr. The reaction mixture was cooled under ice
bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (50 mL.times.2). The
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford crude which was purified by column
chromatography using silica gel (100-200 mesh) and 5% EtOAc/hexane
as eluent to afford 300 mg of
2-ethyl-3-hydroxypyridine-4-carbaldehyde. GCMS: 151 (m/z).
Step 5:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-2-ethyl-pyridin-3-ol
##STR00189##
[0519] 2-Ethyl-3-hydroxy-pyridine-4-carbaldehyde (200 mg, 1.3245
mmol) was taken in mixed solvent [TFE (2 mL): MeCN (2 mL)] and
4-fluoro-3-chloro phenyl amine (192 mg, 1.3245 mmol) was added at
25.degree. C. The resulting mixture was stirred at this temperature
for 2 hr. The reaction mass was concentrated to afford 350 mg of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-2-ethyl-pyridin-3-ol
(crude). LCMS: 279 (M+H).
Step 6:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-ethyl-furo[2,3-c]pyridine-2,3-
-diamine
##STR00190##
[0521] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-2-ethyl-pyridin-3-ol
(crude) (300 mg, 1.0701 mmol) in mixed solvent [DCM (2 mL): TFE (2
mL)] was added TMSCN (0.5 mL, 3.777 mmol) at 25.degree. C. The
reaction mixture was stirred for 2 hr at 25.degree. C.,
concentrated, and the crude material was purified by column
chromatography to yield 80 mg (24% yield) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-ethyl-furo[2,3-c]pyridine-2,3-diamin-
e as a light brown solid. HPLC: 95.02%, LCMS: 306 (M+H),
.sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta. 7.97 (d, 1H, J=5.1
Hz), 7.14-7.08 (m, 2H), 6.81 (s, 2H), 6.72 (d, 1H, J=5.1 Hz),
6.55-6.53 (m, 1H), 6.48-6.44 (m, 1H), 2.88-2.83 (q, 2H), 1.28 (t,
3H, J=7.6 Hz).
Example 10
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-propyl-furo[2,3-c]pyridine-2,3-diami-
ne (Compound 77)
##STR00191##
[0522] Step 1: 2-Chloro-3-methoxymethoxypyridine
##STR00192##
[0524] To a stirred solution of 2-chloro-3-hydroxypyridine (5 g,
38.59 mmol) in THF: DMF (10:25 mL) at 0.degree. C. was added t-BuOK
(4.763 g, 42.45 mmol) portionwise. After stirring the reaction
mixture for 15 min, methoxymethyl chloride (3.062 mL, 40.5 mmol)
was added at 0.degree. C. and the resulting mixture was stirred for
1 hr at 25.degree. C. The reaction mixture was diluted with water
and extracted with ethyl acetate (3.times.150 mL). The combined
organic layer was dried over anhydrous sodium sulfate, concentrated
under reduced pressure to afford crude which was purified by column
chromatography using silica (100-200 mesh) and 10% EtOAc-hexane as
eluent to afford 2-chloro-3-methoxymethoxypyridine (5.4 g) as a
pale brown liquid. LCMS: 174 (M+H).
Step 2: 3-Methoxymethoxy-2-propyl-pyridine
##STR00193##
[0526] To a stirred solution of 2-chloro-3-methoxymethoxypyridine
(500 mg, 2.89 mmol) in Et.sub.2O (5 mL) was added Ni(dppp)Cl.sub.2
(15 mg, 0.028 mmol). The reaction mixture was cooled to 0.degree.
C., propylmagnesium chloride (2 M, 4.3 mL, 8.6 mmol) was added and
resulting mixture was refluxed for 3 hr. The reaction mass was
allowed to come to rt (25.degree. C.), acidified with 2 N HCl, and
washed with MTBE. The aqueous part was basified with solid
K.sub.2CO.sub.3, extracted with EtOAc (50 mL.times.2). The combined
organic part was washed with water, followed by brine, dried over
Na.sub.2SO.sub.4, and evaporated to dryness to provide crude
3-methoxymethoxy-2-propylpyridine (430 mg). LCMS: 182 (M+H).
Step 3: 3-Methoxymethoxy-2-propyl-pyridine-4-carbaldehyde
##STR00194##
[0528] To a stirred solution of 3-methoxymethoxy-2-propylpyridine
(650 mg, 3.591 mmol) in anhydrous THF (6 mL) was added TMEDA (0.6
mL, 3.95 mmol) at 25.degree. C. The reaction mixture was cooled to
-78.degree. C. and n-SuLi (2.17 M in hexane; 1.82 mL, 3.95 mmol)
was added dropwise maintaining the temperature at -78.degree. C.
After stirring for 2 hr at -78.degree. C., DMF (0.4 mL, 5.207 mmol)
was added, the reaction stirred for 1 hr at -78.degree. C., and the
temperature slowly raised to 25.degree. C. The reaction mixture was
quenched with saturated ammonium chloride solution, extracted with
ethyl acetate (50 mL.times.2), the EtOAc part was washed with water
followed by brine, dried over sodium sulfate and concentrated under
reduced pressure to afford crude material which was purified by
column chromatography to afford 480 mg of
3-methoxymethoxy-2-propyl-pyridine-4-carbaldehyde as a pale yellow
liquid. GCMS: 209 (m/z).
Step 4: 3-Hydroxy-2-propyl-pyridine-4-carbaldehyde
##STR00195##
[0530] To a stirred solution of
methoxymethoxy-2-propylpyridine-4-carbaldehyde (450 mg, 2.153 mmol)
in THF (5 mL) was added 3N HCl (6 mL) and the mixture stirred at
60.degree. C. for 2 hr. The reaction mixture was cooled under ice
bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (25 mL.times.2). The
organic layer was dried over anhydrous sodium sulfate, concentrated
under reduced pressure to afford crude product which was purified
by column chromatography using silica gel (100-200 mesh) and
EtOAc/hexane as eluent to afford 285 mg of
3-hydroxy-2-propyl-pyridine-4-carbaldehyde. .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.34 (bs, 1H), 10.24 (s, 1H),
8.17(d, 1H), 4.43 (d, 1H), 2.80 (t, 2H), 1.69 (m, 2H), 0.92 (t,
3H).
Step 5:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-2-propyl-pyridin-3-ol
##STR00196##
[0532] 3-Hydroxy-2-propylpyridine-4-carbaldehyde (274 mg, 1.66
mmol) was taken in mixed solvent [TFE (2 mL):MeCN(2 mL)],
4-fluoro-3-chlorophenyl amine (265 mg, 1.82 mmol) was added at
25.degree. C., and the resulting mixture was stirred at this
temperature for 2 hr. The reaction mass was concentrated to afford
520 mg of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-2-propylpyridin-3-ol.
LCMS: 292.8 (M+H).
Step 6:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-propyl-furo[2,3-c]pyridine-2,-
3-diamine
##STR00197##
[0534] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-2-propyl-pyridin-3-ol
(492 mg, 1.684 mmol) in mixed solvent [DCM (4 mL): TFE (4 mL)] was
added TMSCN (1.09 mL, 8.76 mmol) at 25.degree. C. The reaction
mixture was stirred for 4 hr at 25.degree. C., concentrated, and
crude material was triturated with MTBE/pentane to provide 120 mg
(22% Yield) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-propyl-furo[2,3-c]pyridine-2,3-diami-
ne as brown solid. HPLC: 96.27%, LCMS: 320 (M+H), .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz): .delta. 7.96 (d, 1H, J=51 Hz), 7.14-7.07
(m, 2H), 6.81 (s, 2H), 6.71 (d, 1H, J=5.1 Hz), 6.55-6.53 (m, 1H),
6.47-6.44 (m, 1H), 2.81 (t, 2H, J=7.4 Hz), 1.79-1.71 (m, 2H), 0.94
(t, 3H, J=7.4 Hz).
Example 11
Synthesis of
N.sup.3-(3-Chloro-4-fluoro-phenyl)-5-methoxy-7-methyl-furo[2,3-c]pyridine-
-2,3-diamine (Compound 85)
##STR00198##
[0535] Step 1: 6-Iodo-2-Methyl-Pyridine-3-ol
##STR00199##
[0537] To a stirred solution of 2-methylpyridine-3-ol (500 mg,
4.5816 mmol) in water (10 mL) was added K.sub.2CO.sub.3 (2.2 g,
16.0359 mmol) and the mixture stirred at room temperature for 10
min. The reaction mass was cooled in an ice bath, iodine (1.4 g,
5.4980 mmol, dissolved in methanol (3 mL)) was added in dropwise
manner and the resulting mixture was allowed to stir at room
temperature for 16 hr. The reaction mass was cooled under ice bath,
sodium thiosulphate solution was added and the mixture stirred for
5 min. The reaction mixture was diluted with water (50 mL), and
extracted with ethyl acetate (4.times.50 mL). The combined organic
part was dried over anhydrous sodium sulfate, and concentrated
under reduced pressure to afford 400 mg crude product.
Precipitation/trituration using DCM:pentane (1:1) afforded
6-iodo-2-methylpyridine-3-ol 200 mg as white solid. LCMS: 234
(M-H).
Step 2: 6-Iodo-3-methoxymethoxy-2-methyl pyridine
##STR00200##
[0539] To a stirred solution of 6-iodo-2-methylpyridine-3-ol (200
mg, 0.851 mmol) in DCM (5 mL) under nitrogen atmosphere at
-78.degree. C. was added methoxymethyl chloride (0.077 mL, 1.0212
mmol) dropwise followed by DIPEA (0.218 mL, 1.2765 mmol) and the
mixture was stirred for 3 hr. The reaction mixture was diluted with
water and extracted with DCM (4.times.20 mL). The combined organic
part was dried over anhydrous sodium sulfate, concentrated under
reduced pressure to afford 250 mg crude which was purified by
column chromatography using silica (100-200 mesh) and 10%
EtOAc-hexane as eluent to afford 6-iodo-3-methoxy methoxy-2-methyl
pyridine (200 mg) as colorless liquid. GCMS: 279 (m/z).
Step 3: 2-Methoxy-3-methoxymethoxy-pyridine-4-carbaldehyde
##STR00201##
[0541] Na metal (86 mg, 3.5842 mmol) was added portion wise to
methanol (5 mL) at 0.degree. C. and the solution stirred for 30
min. 6-Iodo-3-methoxymethoxy-2-methylpyridine (200 mg, 0.7168 mmol)
in methanol (5 mL) was added to the reaction mixture followed by
CuBr (20.5 mg, 0.1433 mmol), and the resulting mixture was refluxed
for 16 hr. The reaction mixture was cooled to room temperature,
filtered through a Celite.RTM. reagent bed and the filtrate was
concentrated. The residue was extracted with EtOAc (20 mL.times.5)
washed with water followed by brine, dried over sodium sulfate and
concentrated under reduced pressure to afford 200 mg of crude which
was purified by column chromatography to afford 80 mg of
6-methoxy-3-methoxymethoxy-2-methylpyridine as colorless liquid.
GCMS: 183 (m/z).
Step 4:
6-Methoxy-3-methoxymethoxy-2-methylpyridine-4-carbaldehyde
##STR00202##
[0543] To a stirred solution of
6-methoxy-3-methoxymethoxy-2-methylpyridine (1 g, 5.4644 mmol) in
anhydrous THF (20 mL) was added TMEDA (0.9 mL, 6.0109 mmol) at
25.degree. C. The reaction mixture was cooled to -78.degree. C.,
and n-BuLi (2.7 mL, 6.0109 mmol, 2.17 M in hexane) was added
dropwise maintaining the temperature at -78.degree. C. After
stirring for 2 hr at -78.degree. C., DMF (0.6 mL, 7.9233 mmol) was
added and the mixture stirred for 2 hr at 25.degree. C. The
reaction mixture was cooled to -40.degree. C. and saturated
ammonium chloride solution was added dropwise. The reaction mass
was extracted with ethyl acetate (100 mL.times.2), the EtOAc part
was washed with water followed by brine, dried over sodium sulfate
and concentrated under reduced pressure to afford 1.5 g of crude
product which was passed through a pad of silica (100-200 mesh)
using 5% EtOAc-hexane as eluent to afford 1 g of crude
6-methoxy-3-methoxymethoxy-2-methylpyridine-4-carbaldehyde as a
pale yellow liquid. GCMS: 211 (m/z).
Step 5: 3-hydroxy-6-methoxy-2-methyl-pyridine-4-carbaldehyde
##STR00203##
[0545] To a stirred solution of
6-methoxy-3-methoxymethoxy-2-methylpyridine-4-carbaldehyde (1g,
4.7393 mmol) in THF (3 mL) was added 3N HCl (10 mL) and the mixture
stirred at 60.degree. C. for 1 hr. The reaction mixture was cooled
under ice bath and pH adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (50 mL.times.5). The
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford 700 mg of crude product which was
purified by column chromatography using silica gel (100-200 mesh)
and 5% EtOAc/hexane as eluent to afford 350 mg of
3-hydroxy-6-methoxy-2-methylpyridine-4-carbaldehyde as pale yellow
solid. GCMS: 167 (m/z).
Step 6:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-6-methoxy-2-methyl-pyri-
din-3-ol
##STR00204##
[0547] To a stirred solution of
3-hydroxy-6-methoxy-2-methylpyridine-4-carbaldehyde (150 mg, 0.8982
mmol) was taken in mixed solvent [TFE (2 mL):MeCN (2 mL)],
4-fluoro-3-chlorophenylamine (130 mg, 0.8982 mmol) was added at
25.degree. C., and the resulting mixture was stirred at this
temperature for 1 hr. The reaction mass was concentrated and
purified by triturating with n-pentane to afford 200 mg of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-6-methoxy-2-methyl-pyridin-3-o-
l as yellow solid. LCMS: 293 (M-H).
Step 7:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-5-methoxy-7-methyl-furo[2,3-c]p-
yridine-2,3-diamine
##STR00205##
[0549] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-6-methoxy-2-methylpyridin-3-ol
(200 mg, 0.6802 mmol) in mixed solvent [DCM (2 mL): TFE (2 mL)] was
added TMSCN (0.297 mL, 2.3809 mmol) at 25.degree. C. The reaction
mixture was stirred 3 hr at 25.degree. C., concentrated, and the
crude material was triturated with n-pentane to yield 80 mg (36%
Yield) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-5-methoxy-7-methyl-furo[2,3-c]pyridine-
-2,3-diamine as a brown solid. HPLC: 98.54%, LCMS: 320 (M-H),
.sup.1H-NMR (DMSO-d.sub.6, 400 MHz): .delta. 7.11 (t, 1H, J=9.04
Hz), 7.01 (bs, 1H), 6.84 (bs, 2H), 6.54-6.51 (m, 1H), 6.47-6.43 (m,
1H), 5.92 (s, 1H), 3.73 (s, 3H), 2.40 (s, 3H).
Example 12
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-7-(pyridin-3-yl)furo[2,3-c]pyridine-2,3-
-diamine (Compound 94)
##STR00206##
[0550] Step 1: Furan-2-ylpyridin-3-ylmethanone
##STR00207##
[0552] To a stirred solution of furan (5 g, 5.37 mL, 73.44 mmol) in
anhydrous THF (80 mL) was added n-BuLi (37.2 mL, 80.79 mmol, 2.17 M
in hexane) by dropwisc maintaining the temperature at -78.degree.
C. After stirring for 20 min at -78.degree. C., 3-cyanopyridine
(7.64 g, 73.44 mmol) in THF (20 mL) was added dropwise and the
mixture stirred for 1 hr at -78.degree. C. Ammonium chloride was
added dropwise to the reaction mixture and acidified with 6N HCl to
pH 3. The reaction mass was extracted with ethyl acetate (150
mL.times.4), the EtOAc part was dried over sodium sulfate and
concentrated under reduced pressure to afford the crude product
which was passed through a pad of silica (100-200 mesh) using 10%
EtOAc-hexane as eluent to afford 5.2 g of
furan-2-ylpyridin-3-yl-methanone as a pale yellow solid. LCMS:
174.1 (M+H).
Step 2: [2,3]Bipyridinyl-3-ol
##STR00208##
[0554] The solution of furan-2-ylpyridin-3-ylmethanone (4.7 g,
27.167 mmol) in CH.sub.3OH (15 mL) and NH.sub.4OH (40 mL) was
charged in an autoclave chamber and allowed to stir for 24 hr at
140.degree. C. The reaction mixture was cooled to 25.degree. C.,
concentrated under reduced pressure, the crude residue was taken in
DCM and washed with 8N NaOH solution (40 mL.times.2). The aqueous
part was neutralized with 6N HCl solution to pH 7 and extracted
with ethyl acetate (100 mL.times.5). The EtOAc part was dried over
sodium sulfate and concentrated under reduced pressure to afford
2.6 g crude [2,3']bipyridinyl-3-ol as a pale yellow solid. LCMS:
173 (M+H).
Step 3: 3-Methoxymethoxy-[2,3']bipyridinyl
##STR00209##
[0556] To a stirred solution of [2,3']bipyridinyl-3-ol (0.5 g,
2.906 mmol) in THF (10 mL) at 0.degree. C. was added t-BuOK (0.358
g, 3.197 mmol) portionwise. After stirring the reaction mixture for
20 min, methoxy methyl chloride (0.23 mL, 3.052 mmol) was added at
0.degree. C. and the resulting mixture was stirred for 2 hr at
25.degree. C. The reaction mass was concentrated under reduced
pressure and the crude residue was taken in 10% IPA/DCM (50 mL) and
basified with NH.sub.4OH solution. The organic part was separated,
dried over anhydrous sodium sulfate, concentrated under reduced
pressure to afford crude which was purified by column
chromatography using silica (100-200 mesh) and 30% EtOAc-hexane as
eluent to afford 3-methoxymethoxy-[2,3']bipyridinyl (0.3 g) as a
pale yellow liquid. LCMS: 217 (M+H).
Step 4: 3-Methoxymethoxy-[2,3]bipyridinyl-4-carbaldehyde
##STR00210##
[0558] To a stirred solution of TMP (0.33 mL, 1.944 mmol) in
anhydrous THF (4 mL) was added n-BuLi (0.85 mL, 1.85 mmol, 2.17 M
in hexane) dropwise maintaining the temperature -30.degree. C. and
the temperature was slowly raised to 0.degree. C. After stirring
the mixture for 30 min at 0.degree. C.,
3-methoxymethoxy-[2,3']bipyridinyl (0.2 g, 0.925 mmol) in anhydrous
THF (2 mL) was added dropwise maintaining the temperature
-78.degree. C. The mixture was stirred for 1 hr at -78.degree. C.,
DMF (0.149 mL, 1.944 mmol) was added and the mixture stirred for 30
min at -78.degree. C. The reaction mixture was cooled to
-40.degree. C. and saturated ammonium chloride solution was added
dropwise. the reaction mass was extracted with ethyl acetate
(5.times.25 mL), the EtOAc part was washed with water followed by
brine, dried over sodium sulfate and concentrated under reduced
pressure to afford 150 mg crude
3-methoxymethoxy-[2,3']bipyridinyl-4-carbaldehyde as pale yellow
liquid. The crude product was used without further
purification.
Step 5: 3-Hydroxy-[2,3']bipyridinyl-4-carbaldehyde
##STR00211##
[0560] To a stirred solution of
3-methoxymethoxy-[2,3']bipyridinyl-4-carbaldehyde (0.15 g, 0.614
mmol) in THF (1 mL) was added 3N HCl (2 mL) and the mixture stirred
at 50.degree. C. for 1 hr. The reaction mixture was cooled in an
ice bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with EtOAc (25 mL.times.5). The
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford crude product which was purified by
column chromatography using silica gel (100-200 mesh) and 5%
methanol in DCM as eluent to afford 70 mg
3-hydroxy-[2,3']bipyridinyl-4-carbaldehyde as a pale yellow solid.
GCMS: 201.2 (M+H).
Step 6:
4-{[3-Chloro-4-fluoro-phenylimino]-methyl}-[2,3]bipyridinyl-3-ol
##STR00212##
[0562] 3-Hydroxy-[2,3']bipyridinyl-4-carbaldehyde (70 mg, 0.35
mmol) was taken in mixed solvent [TFE (1 mL): McCN (1 mL)],
4-fluoro-3-chloro phenyl amine (50 mg, 0.35 mmol) was added at
25.degree. C., and the resulting mixture stirred at this
temperature for 2 hr. The reaction mass was concentrated and
purified by triturating with n-pentane to afford 50 mg of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-[2,3']bipyridinyl-3-ol
as a yellow solid. LCMS: 328 (M+H).
Step 7:
N.sup.3-(3-Chloro-4-fluoro-phenyl)-7-pyridin-3-yl-furo[2,3-c]pyrid-
ine-2,3-diamine
##STR00213##
[0564] To a stirred solution of
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-[2,3']bipyridinyl-3-ol
(50 mg, 0.1529 mmol) in mixed solvent [DCM (1 mL):TFE (1 mL)] was
added TMSCN (0.065 mL, 0.519 mmol) at 25.degree. C. The reaction
mixture was stirred 3 hr at 25.degree. C., and concentrated, crude
material was triturated with n-pentane and CH.sub.3CN to yield 25
mg (Yield: 46%) of
N.sup.3-(3-chloro-4-fluoro-phenyl)-7-pyridin-3-yl-furo[2,3-c]pyridine-2,3-
-diamine as brown solid. HPLC: 99.46%. LCMS: 355 (M+H). .sup.1H-NMR
(CD.sub.3CN, 400 MHz): .delta. 9.51 (s, 1H), 8.63 (m, 2H), 8.25 (m,
1H), 7.51 (m, 1H), 7.03-6.99 (m, 2H), 6.66-6.59 (m, 2H), 5.73 (s,
1H), 5.51 (s, 2H).
Example 13
Synthesis of
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridine-7-carbonitr-
ile (Compound 106)
##STR00214##
[0565] Step 1: 3-(Methoxymethoxy)picolinonitrile
##STR00215##
[0567] To a stirred solution of 2-cyano-3-hydroxypyridine (500 mg,
4.162 mmol, 1 eq) in THF:DMF (1:3; 2 mL/mmol) at 0.degree. C. was
added .sup.tBuOK (510 mg, 4.579 mmol, 1.1 eq) portionwise. After
stirring the reaction mixture for 15 min, methoxymethylchloride
(0.33 mL, 4.37 mmol, 1.05 eq) was added at 0.degree. C. and the
resulting mixture was stirred for 1 hr at 25.degree. C. The
reaction mixture was diluted with water and extracted with ethyl
acetate (3.times.30 mL). The organic layer was dried over anhydrous
sodium sulfate, concentrated under reduced pressure to afford the
crude mixture, which was purified by column chromatography to
afford 400 mg of the product. Yield=58%. LCMS: 165.4 (M+H).
Step 2: 4-Formyl-3-(methoxymethoxy)picolinonitrile
##STR00216##
[0569] To a stirred solution of TMP (0.43 mL, 2.50 mmol, 2.05 eq)
in anhydrous THF (3 mL/mmol) was added n-BuLi (1.12 mL, 2.43 mmol,
2 eq, 2.17 M in hexane) dropwise at -40.degree. C. After stirring
for 10 min at -40.degree. C., the reaction mixture was warmed to
0.degree. C. and stirred for 20 min. The reaction mixture was
cooled to -78.degree. C. and 3-(methoxymethoxy)picolinonitrile (200
mg, 1.219 mmol, 1 eq) in THF (2.5 mL/mmol) was added slowly. After
stirring for 30 min at -78.degree. C., DMF (0.19 mL, 2.50 mmol,
2.05 eq) was added and stirred for another 30 min at -78.degree. C.
Saturated ammonium chloride solution was then added dropwise below
-50.degree. C. and the reaction mass was cooled to 0.degree. C. The
reaction mass was extracted with ethyl acetate (3.times.30 mL). The
organic layer was washed with water followed by brine, dried over
sodium sulfate and concentrated under reduced pressure to afford
the crude product which was used for next step without
purification.
Step 3: 4-Formyl-3-hydroxypicolinonitrile
##STR00217##
[0571] To a stirred solution of crude
4-formyl-3-(methoxymethoxy)picolinonitrile (200 mg, 1 eq) in THF (3
mL/mmol) was added 3N HCl (2 mL) and the mixture stirred at
60.degree. C. for 1 hr. The reaction mixture was cooled under ice
bath and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The
resulting mixture was extracted with ethyl acetate (2.times.50 mL).
The organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford the crude which was purified by column
chromatography to afford 80 mg of crude product.
Step 4:
(E)-4-(((3-Chloro-4-fluorophenyl)imino)methyl)-3-hydroxypicolinoni-
trile
##STR00218##
[0573] This compound was prepared according to Procedure-A and was
carried forward to next step as such without further
purification.
Step 5:
2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridine-7-ca-
rbonitrile
##STR00219##
[0575] Following general procedure A,
2-amino-3-((3-chloro-4-fluorophenyl)amino)
furo[2,3-c]pyridine-7-carbonitrile was prepared (7 mg). HPLC:
95.29%; LCMS: 303 (M+H); .sup.1H-NMR (CD.sub.3CN, 400 MHz): .delta.
8.19 (d, 1H), 7.15 (d, 1H), 7.03 (t, 1H), 6.63-6.61 (m, 1H),
6.56-6.52 (m, 1H), 5.81 (s, 2H), 5.71 (s, 1H).
Example 14
Synthesis of
N.sup.3-(3-Chloro-4-fluorophenyl)-5-phenoxy-7-phenylfuro[2,3-c]pyridine-2-
,3-diamine (Compound 104)
##STR00220##
[0576] Step 1: 2-Phenylpyridin-3-ol
##STR00221##
[0578] To a stirred solution of 2-iodo-3-hydroxypyridin (5.0 g,
22.624 mmol) in benzene (50 ml) was added phenylboronic acid (3.03
g, 24.85 mmol) and 2M Na.sub.2CO.sub.3 solution (20 mL) at
25.degree. C. and the reaction mixture was degassed with argon for
15 min. Then added Pd(PPh.sub.3).sub.4 (1.3 g, 5 mol %) and further
degassed for 10 min. The resulting reaction mixture was refluxed
for 4 h. After completion of reaction the reaction mixture was
cooled to room temperature, water was added to it and extracted
with EtOAc. Organic layer was dried over anhydrous sodium sulphate,
concentrated under reduced pressure to afford the crude which was
purified by trituration with MTBE/DCM to afford
2-phenylpyridin-3-ol (3.2 g). LCMS: 172 (M+H).
Step 2: 6-Iodo-2-phenylpyridin-3-ol
##STR00222##
[0580] To a stirred solution of 2-phenylpyridin-3-ol (3.0 g, 17.524
mmol) in THF (75 mL) was added Na.sub.2CO.sub.3 (3.9 g, 36.792
mmol) in 75 ml water and stirred at room temperature for 10 min.
Reaction mass was cooled under ice bath, then iodine (4.45 g,
35.066 mmol) was added portion wise and the resulting mixture was
allowed to stir at room temperature for 16 h. Reaction mass was
cooled under ice-bath and sodium thiosulphate solution was added to
it and stirred for 5 min. Reaction mixture was diluted with water,
extracted with ethyl acetate. Combined organic part was dried over
anhydrous sodium sulphate, concentrated under reduced pressure to
afford the crude material which was purified by column
chromatography to afforded 6-iodo-2-phenylpyridin-3-ol (1.6 g).
LCMS: 297.8 (M+H).
Step 3: 6-Iodo-3-(methoxymethoxy)-2-phenylpyridine
##STR00223##
[0582] To a stirred solution of 6-iodo-2-phenylpyridin-3-ol (3.3 g,
11.107 mmol) in a mixture of solvents THF (6 mL):DMF (15 mL) at
0.degree. C. was added t-BuOK (1.5 g, 13.368 mmol) portion wise.
After stirring the reaction mixture for 15 mins, methoxymethyl
chloride (0.98 mL, 12.173 mmol) was added to it at 0.degree. C. and
the resulting mixture was stirred for 1 h at 25.degree. C. Reaction
mixture was diluted with water and extracted with ethyl acetate.
Organic layer was dried over anhydrous sodium sulphate,
concentrated under reduced pressure to afford the crude mass which
was purified by column chromatography to afford
6-iodo-3-(methoxymethoxy)-2-phenylpyridine (2.4 g). LCMS: 341.8
(M+H).
Step 4: 3-(Methoxymethoxy)-6-phenoxy-2-phenylpyridine
##STR00224##
[0584] Na metal (0.38 g, 17.272 mmol) was added portionwise to
phenol (20 mL) at 0.degree. C. and stirred for 30 min at room
temperature, then 6-iodo-3-(methoxymethoxy)-2-phenyl pyridine (1.2
g, 3.517 mmol) was added followed by CuBr (0.096 g, 0.669 mmol),
the resulting reaction mixture was refluxed for 16 h and after
completion of reaction the reaction mixture was cooled to room
temperature and filtered through celite bed and the filtrate was
concentrated. Residue was extracted with EtOAc, washed with water
followed by brine, dried over sodium sulphate and concentrated
under reduced pressure to afford the crude material which was
purified by column chromatography to get
3-(methoxymethoxy)-6-phenoxy-2-phenylpyridine (0.8 g). LCMS: 308
(M+H).
Step 5: 3-(Methoxymethoxy)-6-phenoxy-2-phenylisonicotinaldehyde
##STR00225##
[0586] To a stirred solution of
3-(methoxymethoxy)-6-phenoxy-2-phenylpyridine (0.2 g, 0.65 mmol) in
anhydrous THF (3 mL) was added TMEDA (0.107 mL, 0.920 mmol) at
25.degree. C. The reaction mixture was cooled to -78.degree. C. and
then added n-BuLi (0.3 mL, 2.2 M) by dropwise at -78.degree. C. and
stirred for 1 h at -78.degree. C. DMF (0.072 mL) was added to it at
-78.degree. C. and the resulting mixture was allowed to stirred for
1 h at -78.degree. C. Saturated ammonium chloride solution was then
added drop wise below -50.degree. C. and the reaction mass was
diluted with ethyl acetate, extracted with ethyl acetate, washed
with brine and dried over sodium sulphate and concentrated to
afford crude
3-(methoxymethoxy)-6-phenoxy-2-phenylisonicotinaldehyde (0.15 g)
which was used in next step without purification. LCMS: 336.2
(M+H).
Step 6: 3-Hydroxy-6-phenoxy-2-phenylisonicotinaldehyde
##STR00226##
[0588] To a stirred solution of crude
3-(methoxymethoxy)-6-phenoxy-2-phenylisonicotinaldehyde (0.15 g,
0.447 mmol) in THF (2 mL) was added 3N HCl (3 mL) and stirred at
60.degree. C. for 1 h. Reaction mixture was cooled under ice-bath
and pH was adjusted to 7 with solid K.sub.2CO.sub.3. The resulting
mixture was extracted with EtOAc, washed with brine, dried over
sodium sulphate and concentrated under reduced pressure to afford
the crude material which was purified by column chromatography to
afford 3-hydroxy-6-phenoxy-2-phenylisonicotinaldehyde (0.055 g).
LCMS: 289.6 (M-H).
Step 7:
N.sup.3-(3-Chloro-4-fluorophenyl)-5-phenoxy-7-phenylfuro[2,3-c]pyr-
idine-2,3-diamine (Procedure C)
##STR00227##
[0590] To a solution of
3-hydroxy-6-phenoxy-2-phenyl-pyridine-4-carbaldehyde (0.050 g,
0.171 mmol) in DCM (2 mL) was added 4-fluoro-3-chloroaniline (0.025
g, 0.171 mmol), TMSOTf (0.006 mL, 0.0343 mmol) and TMSCN (0.11 mL,
0.893 mmol). The resulting mixture was stirred for 6 h at room
temperature. After completion of reaction the reaction mass was
diluted with DCM, washed with water followed by brine and dried
over anhydrous Na.sub.2SO.sub.4 and was evaporated to provide a
brown crude material which was further purified by column
chromatography/trituration to afford
N.sup.3-(3-chloro-4-fluoro-phenyl)-5-phenoxy-7-phenyl-furo[2,3-c]pyridine-
-2,3-diamine (0.015 g, 0.033 mmol, yield 20%). .sup.1H-NMR (400
MHz, DMSO-d.sub.6): .delta. 8.11 (d, J=6.6 Hz, 2H), 7.47-7.37 (m,
5H), 7.15-7.11 (m, 7H), 6.60 (bs, 1H), 6.52 (bs, 1H), 6.26 (s, 1H);
LCMS: 444.2 (M+H).
Example 15
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-5-ethyl-7-phenylfuro[2,3-c]pyridine-2,3-
-diamine (Compound 109)
##STR00228##
[0591] Step 1: 2-Phenylpyridin-3-ol
##STR00229##
[0593] To a stirred solution of 2-iodo-3-hydroxypyridine, prepared
as described above, (5 g, 22.62 mmol, 1 eq) in DME (2 mL/mmol) was
added phenylboronic acid (3.03 g, 24.88 mmol, 1.1 eq) and 2M
Na.sub.2CO.sub.3 solution (20 mL, 1.7 eq) at 25.degree. C. and the
reaction mixture was degassed with argon for 15 min.
Pd(PPh.sub.3).sub.4 (1.3 g, 5 mol %) was then added, the mixture
was further degassed for 10 min, and the reaction mixture refluxed
for 4 hr. The reaction mixture was cooled to RT, filtered through a
pad of Celite.RTM. reagent and the filtrate was concentrated. The
residue was diluted with ethyl acetate (2.times.50 mL) and washed
with water and brine. The organic layer was dried over anhydrous
sodium sulfate, concentrated under reduced pressure to afford the
crude which was purified by column chromatography to afford 1.6 g
of 2-phenylpyridin-3-ol (Yield: 41%). LCMS: 172 (M+H).
Step 2: 6-Iodo-2-phenylpyridin-3-ol
##STR00230##
[0595] To a stirred solution of 2-phenylpyridin-3-ol (3 g, 17.54
mmol, 1 eq) in THF (4 mL/mmol) was added Na.sub.2CO.sub.3 (3.9 g,
36.83 mmol, 2.1 eq; 0.5 M in water) and stirred at room temperature
for 10 min. The reaction mass was cooled under ice bath. Iodine
(4.45 g, 17.54 mmol, 1 eq) was added portionwise and the resulting
mixture was allowed to stir at room temperature for 16 hr. The
reaction mass was cooled under ice bath, sodium thiosulphate
solution was added to it and stirred for 5 min. The reaction
mixture was diluted with water, and extracted with ethyl acetate.
The combined organic part was dried over anhydrous sodium sulfate,
and concentrated under reduced pressure to afford the crude
product. Purification of the crude material by column
chromatography afforded 1.6 g of 6-iodo-2-phenylpyridin-3-ol
(Yield: 31%). LCMS: 297.8 (M+H).
Step 3: 6-Iodo-3-(methoxymethoxy)-2-phenylpyridine
##STR00231##
[0597] To a stirred solution of 6-iodo-2-phenylpyridin-3-ol (3.3 g,
11.11 mmol, 1 eq) in THF:DMF (1:3, 2 mL/mmol) at 0.degree. C. was
added t-BuOK (1.5 g, 13.33 mmol, 1.2 eq) portionwise. After
stirring the reaction mixture for 15 min, methoxymethylchloride
(0.91 mL, 12.17 mL, 1.1 eq) was added at 0.degree. C. and the
resulting mixture was stirred for 1 hr at 25.degree. C. The
reaction mixture was diluted with water and extracted with ethyl
acetate (2.times.100 mL). The organic layer was dried over
anhydrous sodium sulfate, concentrated under reduced pressure to
afford the crude product, which was purified by column
chromatography to afford 2.4 g of
6-oodo-3-(methoxymethoxy)-2-phenylpyridine (Yield: 63%). LCMS:
341.8 (M+H).
Step 4: 6-Ethyl-3-(methoxymethoxy)-2-phenylpyridine
##STR00232##
[0599] To a stirred solution of
6-iodo-3-(methoxymethoxy)-2-phenylpyridine (900 mg, 2.63 mmol, 1
eq) in DMF (10 mL/mmol) was added triethylborate (5.27 mL, 5.27
mmol, 1M in THF, 2 eq) and K.sub.2CO.sub.3 (546 mg, 3.95 mmol, 1.5
eq) at 0.degree. C. The reaction mixture was degassed with argon
and [1,3-bis(diphenylphosphino)propane]dichloronickel (II) (76 mg,
0.0659 mmol, 2.5 mol %) was added. The reaction mixture was heated
at 80.degree. C. for 16 hr. The reaction mass was cooled to room
temperature, filtered through Celite.RTM. reagent, filtrate was
diluted with water and acidified (pH 4) using 1N HCl and stirred
for 15 min. To this suspension saturated NaHCO.sub.3 solution was
added to make the pH 9, extracted with MTBE, organic part was dried
over Na.sub.2SO.sub.4, evaporated to dryness and the crude product
was purified by column chromatography to yield 500 mg of
6-ethyl-3-(methoxymethoxy)-2-phenylpyridine (Yield: 77%). LCMS:
243.8 (M+H).
Step 5: 6-Ethyl-3-(methoxymethoxy)-2-phenylisonicotinaldehyde
##STR00233##
[0601] To a stirred solution of
6-ethyl-3-(methoxymethoxy)-2-phenylpyridine (200 mg, 0.8230 mmol, 1
eq) in anhydrous THF (3 mL/mmol) was added TMEDA (0.135 mL, 0.9053
mmol, 1.1 eq) at 25.degree. C. The reaction mixture was cooled to
-78.degree. C., n-BuLi (0.417 mL, 0.9053 mmol, 2.17M, 1.1 eq) was
added by dropwise at -78.degree. C. and stirred for 1 hr at
-78.degree. C. DMF (0.091 mL, 1.1934 mmol, 1.45 eq) was added to it
at -78.degree. C. and the resulting mixture was allowed to stir for
1 hr at -78.degree. C. Saturated ammonium chloride solution was
then added dropwise below -50.degree. C. and the reaction mass was
diluted with ethyl acetate. The organic layer was separated, washed
with brine and dried over sodium sulfate. The organic layer was
filtered and concentrated to afford 200 mg of crude
6-ethyl-3-(methoxymethoxy)-2-phenylisonicotinaldehyde which was
used for next step without purification. LCMS: 271.8 (M+H).
Step 6: 6-Ethyl-3-hydroxy-2-phenylisonicotinaldehyde
##STR00234##
[0603] To a stirred solution of crude
6-ethyl-3-(methoxymethoxy)-2-phenylisonicotinaldehyde (200 mg,
0.7380 mmol, 1 eq) in THF (3 mL/mmol) was added 3N HCl (6 mL/mmol)
and stirred at 60.degree. C. for 1 hr. The reaction mixture was
cooled under ice bath and pH was adjusted to 7 with solid
K.sub.2CO.sub.3. The resulting mixture was extracted with ethyl
acetate (2.times.50 mL). The organic layer was dried over sodium
sulfate and concentrated under reduced pressure to afford the crude
product which was purified by column chromatography to afford 120
mg of 6-ethyl-3-hydroxy-2-phenylisonicotinaldehyde (Yield: 64%
after two steps). LCMS: 226.2 (M-H).
Step 7:
(E)-4-(((3-Chloro-4-fluorophenyl)imino)methyl)-6-ethyl-2-phenylpyr-
idin-3-ol
##STR00235##
[0605]
(E)-4-(((3-Chloro-4-fluorophenyl)imino)methyl)-6-ethyl-2-phenylpyri-
din-3-ol (50 mg) was prepared by following general procedure A.
LCMS: 355.0 (M+H).
Step 8:
N3-(3-Chloro-4-fluorophenyl)-5-ethyl-7-phenylfuro[2,3-c]pyridine-2-
,3-diamine
##STR00236##
[0607]
N.sup.1-(3-chloro-4-fluorophenyl)-5-ethyl-7-phenylfuro[2,3-c]pyridi-
ne-2,3-diamine (15 mg) was prepared by following general procedure
A. HPLC: 97.61%; LCMS: 382 (M+H); .sup.1H-NMR (CDCl.sub.3, 400
MHz): .delta. 8.25 (d, 2H), 7.50 (t, 2H), 7.42 (m, 1H), 6.96 (t,
1H), 6.81 (s, 1H), 6.65 (m, 1H), 6.51 (m, 1H), 4.74 (s, 1H), 4.49
(s, 2H), 2.84 (q, 2H), 1.32 (t, 3H).
Example 16
Synthesis of
2-Amino-3-((3-chloro-4-fluorophenyl)amino)benzofuran-6-carbonitrile
(Compound 112) (Procedure A, B)
##STR00237##
[0609] To a stirred solution of 4-formyl-3-hydroxybenzonitrile (0.2
g, 1.36 mmol) in DCM (4 mL) was added 3-chloro-4-fluoroaniline
(0.198 g, 1.36 mmol) followed by the addition of TMSCN (0.89 mL,
7.074 mmol) and TMS-OTf (0.049 mL, 0.272 mmol). The resulting
mixture was stirred in a sealed tube at 25.degree. C. for 16 hr.
After completion of reaction the reaction mixture was diluted with
diethyl ether, washed with water, dried over sodium sulfate and
evaporated under reduced pressure to give crude material obtained
was purified by column chromatography over alumina using
acetone/hexane as eluent to afford
2-amino-3-(3-chloro-4-fluoro-phenylamino)-benzofuran-6-carbonitrile
(0.03 g, 0.103 mmol, 8%) as reddish brown solid. .sup.1H-NMR (400
MHz, DMSO-d.sub.6): .delta. 7.75 (s, 1H), 7.39 (d, J=8.1 Hz, 1H),
7.14-7.10 (m, 2H), 6.96 (bs, 2H), 6.92 (d, J=8.0 Hz, 1H), 6.56 (dd,
J'=6.3 Hz, J''=2.7 Hz, 1H), 6.47 (dt, J'=8.8 Hz, J''=6.6 Hz,
J'''=3.2 Hz, 1H); LCMS: 300 (M-H).
Example 17
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)thieno[2,3-c]pyridine-2,3-diamine
(Compound 121)
##STR00238##
[0610] Step 1: 3-Chloropyridine-4-carbonitrile
##STR00239##
[0612] To a stirred solution of 2,2,6,6-tetramethyl piperidine
(17.16 mL, 100.854 mmol) in THF (100 mL) was added n-BuLi (2.17 M,
44.26 mL, 96.052 mmol) at -30.degree. C. After stirring the mixture
for 15 min at 25.degree. C., it was cooled to -78.degree. C. and
4-cyanopyridine (5 g, 48.026 mmol) in THF (40 mL) was added
dropwise. After stirring the reaction mixture for 30 min at
-78.degree. C., hexachloroethane (23.87 g, 100.854 mmol) in THF (50
mL) was added at -78.degree. C. The resulting mixture was stirred
for 30 min at -78.degree. C. and was quenched with saturated
NH.sub.4Cl solution. Water (100 mL) was added to the reaction
mixture and extracted with ethyl acetate (4.times.300 mL). The
combined organic layer was dried over anhydrous sodium sulfate,
concentrated under reduced pressure to afford the crude material
which was purified by column chromatography using silica (100-200
mesh) and 5% EtOAc-hexane as eluent to afford
3-chloropyridine-4-carbonitrile (3.5 g, 25.261 mmol, 53%) as pale
white solid. GCMS: 138 (m/z).
Step 2: Methyl 3-aminothieno[2,3-c]pyridine-2-earboxylate
##STR00240##
[0614] To a stirred solution of 3-chloropyridine-4-carbonitrile
(6.2 g, 44.92 mmol) in MeCN (60 mL) was added mercapto-acetic acid
methyl ester (4.26 mL, 47.173 mmol) and K.sub.2CO.sub.3 (12.4 g,
89.855 mmol) at 25.degree. C. The reaction mixture was refluxed for
3 hr and was concentrated under reduced pressure. The resulting
residue was diluted with water (100 mL) and extracted with EtOAc
(4.times.200 mL). The combined organic layer was dried over sodium
sulfate and was concentrated under reduced pressure to afford the
crude material which was purified by trituration with n-pentane to
afford methyl 3-aminothieno[2,3-c]pyridine-2-carboxylate (7.5 g,
36.016 mmol, 80%) as pale yellow solid. LCMS: 209 (M+H).
Step 3: Methyl 3-bromothieno[2,3-c]pyridine-2-carboxylate
##STR00241##
[0616] To a stirred solution of CuBr (2.71 g, 18.930 mmol) in
aqueous HBr (46 mL, 48%) was added methyl
3-aminothieno[2,3-c]pyridine-2-carboxylate (3.75 g, 18.028 mmol) at
-10.degree. C. NaNO.sub.2 (1.49 g, 21.634 mmol) in water (43 mL)
was added dropwise at -10.degree. C. and the resulting mixture was
stirred for 30 min at -10.degree. C. Solid NaNO.sub.2 (0.149 g,
2.163 mmol) was then added to the reaction mass at -10.degree. C.
After stirring for 30 min at -10.degree. C., another portion of
solid NaNO.sub.2 (0.149 g, 2.163 mmol) was added. The reaction
mixture was then slowly poured into saturated NaHCO.sub.3 solution
(pH=7) and extracted with DCM (4.times.200 mL). The combined
organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford crude material which was purified by
column chromatography using silica gel (100-200 mesh) and 5%
EtOAc/hexane as eluent to afford methyl
3-bromothieno[2,3-c]pyridine-2-carboxylate (1.5 g, 5.512 mmol, 31%)
as off white solid. LCMS: 271.8 (M+H).
Step 4: Methyl
3-((3-chloro-4-fluorophenyl)amino)thieno[2,3-c]pyridine-2-carboxylate
##STR00242##
[0618] To a stirred solution of methyl
3-bromothieno[2,3-c]pyridine-2-carboxylate (0.2 g, 0.735 mmol) in
toluene (5 mL) was added 3-chloro-4-fluoro aniline (0.149 g, 1.029
mmol), Cs.sub.2CO.sub.3 (0.337 g, 1.102 mmol), BINAP (0.091g, 0.147
mmol) and degassed for 20 min. Pd.sub.2(dba).sub.3 (0.038 g, 0.036
mmol) was then added and the reaction mixture was heated at
100.degree. C. for 16 hr. The reaction mixture was filtered through
a bed of Celite.RTM. reagent and washed with ethyl acetate. The
filtrate was diluted with water (50 mL) and extracted with EtOAc
(4.times.50 mL). The combined organic layer was dried over sodium
sulfate, concentrated under reduced pressure to afford crude
material which was purified by column chromatography using silica
gel (100-200 mesh) and 5% EtOAc/hexane as eluent to afford methyl
3-((3-chloro-4-fluorophenyl)-amino)thieno[2,3-c]pyridine-2-carboxylate
(0.1 g, 0.296 mmol, 40%) as pale yellow solid. LCMS: 336.8
(M+H).
Step 5: Methyl 3-((tert-butoxycarbonyl)(3-chloro-4
fluorophenyl)amino)thieno-[2,3-e]pyridine-2-carboxylate
##STR00243##
[0620] To a stirred solution of methyl
3-((3-chloro-4-fluorophenyl)amino)thieno[2,3-c]pyridine-2-carboxylate
(0.9 g, 2.678 mmol) in THF(10 mL) was added TEA (0.93 mL, 6.696
mmol) and DMAP (0.196 g, 1.607 mmol) at 0.degree. C. The resulting
mixture was stirred for 10 min at 0.degree. C. and then Boc
anhydride (1.31 mL, 5.892 mmol) was added dropwise. The reaction
mixture was heated at 60.degree. C. for 4 hr. After completion of
the reaction, water (100 mL) was added to the reaction mixture and
extracted with EtOAc (4.times.70 mL). The combined organic layer
was dried over sodium sulfate, concentrated under reduced pressure
to afford the crude material which was purified by column
chromatography using silica gel (100-200 mesh) and 15% EtOAc/hexane
as eluent to afford methyl
3-((tert-butoxycarbonyl)(3-chloro-4-fluorophenyl)amino)-thieno[2,3-c]pyri-
dine-2 carboxylate (0.8 g, 1.831 mmol, 69%). LCMS: 436.8 (M+H).
Step 6:
3-((teat-Butoxycarbonyl)(3-chloro-4-fluorophenyl)amino)thieno[2,3--
c]pyridine-2-carboxylic acid
##STR00244##
[0622] To a stirred solution of methyl
3-((tert-butoxycarbonyl)(3-chloro-4-fluorophenyl)-amino)thieno[2,3-c]pyri-
dine-2 carboxylate (0.8 g, 1.831 mmol) in a mixture of
THF:MeOH:H.sub.2O (15:9:3 mL) was added LiOH (0.088 g, 3.669 mmol)
at 0.degree. C. and resulting mixture was stirred for 16 hr. The
reaction mixture was concentrated under reduced pressure and the
resulting residue was diluted with water (5 mL) and acidified with
5% citric acid solution (pH=4). The resulting solid precipitate was
filtered, washed with water and dried under vacuum to afford
3-((tert-butoxycarbonyl)(3-chloro-4-fluorophenyl)amino)thieno[2,3-c]pyrid-
ine-2-carboxylic acid (0.7 g, 1.655 mmol, 90%) as pale yellow
solid. LCMS: 423 (M.sup.t).
Step 7: teat-Butyl
(2-((tert-butoxycarbonyl)amino)thieno[2,3-c]pyridin-3-yl)(3-chloro-4-fluo-
rophenyl)carbamate
##STR00245##
[0624] To a stirred solution of
3-((tent-butoxycarbonyl)(3-chloro-4-fluorophenyl)
amino)thieno[2,3-c]pyridine-2-carboxylic acid (0.3 g, 0.710 mmol)
in tert-BuOH (10 mL) was added DPPA (0.18 mL, 0.853 mmol) and DIPEA
(0.13 mL, 0.782 mmol) at 25.degree. C. The resulting reaction
mixture was heated to 100.degree. C. for 2 hr and, after completion
of the reaction, the mixture was concentrated under reduced
pressure to give a residue which was diluted with water (30 mL),
neutralized with saturated NaHCO.sub.3 solution and extracted with
EtOAc (4.times.70 mL). The combined organic layer was dried over
sodium sulfate, concentrated under reduced pressure to afford the
crude material which was purified by column chromatography using
silica gel (100-200 mesh) and 2% MeOH/DCM as eluent to afford
tent-butyl
(2-((tert-butoxycarbonyl)amino)thieno[2,3-c]pyridin-3-yl)(3-chloro-4-fluo-
rophenyl)carbamate (0.15 g, 0.303 mmol, 43%) as off white solid.
LCMS: 494 (M+H).
Step 8:
N.sup.3-(3-Chloro-4-fluorophenyl)thieno[2,3-c]pyridine-2,3-diamine
##STR00246##
[0626] To a stirred solution of tent-butyl
(2-((tert-butoxycarbonyl)amino)thieno[2,3-c]pyridin-3-yl)(3-chloro-4-fluo-
rophenyl)carbamate (0.13 g, 0.263 mmol) in 1,4-dioxane (2 mL) was
added 4 M dioxane.HCl solution (0.59 mL, 2.373 mmol) at 0.degree.
C. The resulting mixture was stirred for 8 hr at 25.degree. C. and,
after completion of the reaction, the reaction mass was
concentrated under reduced pressure to give residue which was
diluted with DCM (70 mL) and basified with liq.NH.sub.3. The DCM
layer was dried over sodium sulfate, concentrated under reduced
pressure to afford crude mass which was purified by column
chromatography using silica gel (100-200 mesh) and 2% MeOH/DCM as
eluent to afford
N.sup.3-(3-chloro-4-fluorophenyl)thieno[2,3-c]pyridine-2,3-diamine
(0.015 g, 0.051 mmol, 20%) as off white solid. .sup.1H-NMR (400
MHz, DMSO-d.sub.6): .delta. 8.66 (s, 1H), 8.13 (d, J=5.4 Hz, 1H),
7.27 (s, 1H), 7.11 (t, J=9.1 Hz, 1H), 6.86 (d, J=5.3 Hz, 1H), 6.72
(bs, 2H), 6.50 (dd, J=6.2 Hz, J''=2.4 Hz, 1H), 6.42-6.40 (m, 1H);
LCMS: 294 (M+H).
Example 18
Synthesis of 3-((3-chlorophenyl)thio)benzo[b]thiophen-2-amine
hydrochloride (Compound125)
##STR00247##
[0627] Step 1: 3-Bromo-1-benzothiophene
##STR00248##
[0629] To a solution of benzo[b]thiophene (10 g, 74.516 mmol) in
chloroform (75 mL) and acetic acid (75 mL), was stepwise added NBS
(16.6 g, 93.268 mmol) for 4 hr at 0.degree. C. and the mixture
allowed to stir at room temperature for 48 hr. The progress of the
reaction was monitored by TLC and, after completion of reaction,
the reaction mass was diluted with chloroform (200 mL) and the
resulting mixture was successively washed with a saturated solution
of sodium thiosulfate (200 mL), sodium carbonate (200 mL) and brine
(150 mL). The extracted organic layer was then dried over sodium
sulfate, filtered and evaporated under reduced pressure. The
resulting red liquid was then filtered of a pad of silica gel,
eluting with hexane to afford 3-bromo-1-benzothiophene (15.87 g,
74.474 mmol, 100%) as yellow oil. The .sup.1H NMR and mass was
confirmed by reported literature.
Step 2: 3-Bromo-2-nitro-1-benzothiophene
##STR00249##
[0631] Fuming nitric acid (8.6 mL) was dropwise added to a mixture
of 3-bromo benzothiophene (3 g, 14.155 mmol) in TFA (7.5 mL) and
DCM (15 mL) at 0.degree. C. The reaction was turned greenish, and a
yellow solid precipitated. To this reaction mixture was added DCM
(10 mL) and the mixture stirred at 0.degree. C. for 30 min. The
reaction mass was poured into ice-water (500 mL) and extracted with
DCM. The combined organic layers were washed with brine, dried over
sodium sulfate and evaporated under reduced pressure to give a
yellow solid. The resulting yellow solid was crystallized with DCM
and hexane mixture to afford 3-bromo-2-nitro-1-benzothiophene (1.5
g, 5.811 mmol, 41%) as a yellow solid. The .sup.1H NMR and mass was
confirmed by reported literature.
Step 3: 3-[(3-Chlorophenyl)sulfanyl]-2-nitro-1-benzothiophene
##STR00250##
[0633] 3-Chlorothiophenol was added to a solution of sodium
hydroxide (0.085 g, 2.139 mmol) in water (2 mL) and dioxane (18 mL)
mixture at 0.degree. C. A solution of
3-bromo-2-nitro-1-benzothiophene (0.5 g, 1.945 mmol) in dioxane (2
mL) was then added dropwise and the reaction mixture stirred for 1
h at the same temperature. After completion of the reaction, the
reaction mass was diluted with water, extracted with ethyl acetate
and washed with brine. The organic layer was dried over sodium
sulfate, concentrated under reduced pressure to give residue which
was purified by column chromatography using 4% ethyl acetate and
hexane mixture on silica gel to afford
3-[(3-chlorophenyl)sulfanyl]-2-nitro-1-benzothiophene (0.375 g,
1.165 mmol, 60%) as yellow solid.
[0634] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.83 (d, J=8.3
Hz, 1H), 7.67 (d, J=8.3 Hz, 1H), 7.58-7.55 (m, 1H), 7.38-7.35 (m,
1H), 7.30-7.29 (m, 1H), 7.22-7.21 (m, 1H), 7.20 (s, 1H), 7.18-7.17
(m, 1H).
Step 4: 3-[(3-Chlorophenyl)sulfanyl]-1-benzothiophen-2-amine
##STR00251##
[0636] To the solution of
3-[(3-chlorophenyl)sulfanyl]-2-nitro-1-benzothiophene (0.3 g, 0.932
mmol) in ethyl acetate (10 mL) was added activated Pd/C (0.02 g,
10%) under hydrogen gas (balloon) at room temperature and the
reaction mass stirred for 12 hr. The reaction mixture was filtered
through a Celite.RTM. reagent bed, washed with ethyl acetate, the
filtrate was distilled off to give a residue. The reside was
purified by column chromatography using 6% ethyl acetate and hexane
mixture on silica gel to afford
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine (0.15 g, 0.514
mmol, 55%) as reddish viscous liquid which was used in next stage
without further purification.
Step 5: 3-[(3-Chlorophenyl)sulfanyl]-1-benzothiophen-2-amine
hydrochloride
##STR00252##
[0638] To the solution of
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine (0.15 g, 0.514
mmol) in diethyl ether (5 mL) was added hydrochloride gas absorbed
diethyl ether solution (10 mL). The reaction mass was stirred at
room temperature for 30 min, a brown solid appeared, the solvent
was decanted and the solid dried under vacuum to afford
3-[(3-chlorophenyl)sulfanyl]-1-benzothiophen-2-amine hydrochloride
(0.1 g, 0.304, 60%) as light brown solid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 7.65 (d, J=7.8 Hz, 1H), 7.24 (t, J=7.9 Hz,
1H), 7.20-7.17 (m, 2H), 7.15-7.12 (m, 1H), 7.05-7.01 (m, 1H), 6.97
(d, J=7.8 Hz, 1H), 6.94-6.93 (m, 1H), 6.45 (bh, 2H); HRMS: 290.9949
(M-H).sup.+.
Example 19
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenylbenzo[b]thiophene-2,3-diamine
(Compound 134)
##STR00253##
[0639] Step 1: 1-Bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene
##STR00254##
[0641] 4-Bromothiophenol (10 g, 52.890 mmol) was dissolved in DMF
(25 mL) and potassium carbonate added, followed by a solution of
bromoacetaldehyde diethyl acetal (8.9 mL, 68.750 mmol) in DMF (25
mL) at 0.degree. C. The reaction mixture was warmed to room
temperature and stirred overnight. The progress of the reaction was
monitored by TLC and, after completion of the reaction, the mixture
was diluted with water and extracted using ethyl acetate. The
combined organic layers were washed with brine, dried over
anhydrous sodium sulfate and solvent was evaporated under reduced
pressure to dryness to give residue which was purified by column
chromatography using hexane as eluent on silica gel to afford
1-bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene (14 g, 52.288 mmol,
99%) as yellow liquid. The .sup.1H NMR and mass was confirmed by
reported literature.
Step 2: 5-Bromo-1-benzothiophene
##STR00255##
[0643] Poly phosphoric acid (28 g) was dissolved in chlorobenzene
(25 mL) at 125.degree. C. and a solution of
1-bromo-4-[(2,2-diethoxyethyl)sulfanyl]benzene (14 g, 55.288 mmol)
in chlorobenzene (28 mL) was added under nitrogen. The reaction
mixture was refluxed at 125.degree. C. overnight. Progress of the
reaction was monitored by TLC and, after completion of the
reaction, the mixture was diluted with water and extracted using
toluene. The combined organic layers were washed with brine, dried
over anhydrous sodium sulfate and solvent was evaporated under
reduced pressure to give residue which was purified by column
chromatography using hexane as eluent on silica gel to afford
5-bromo-1-benzothiophene (5 g, 45.867 mmol, 51%) as white solid.
The .sup.1H NMR and mass was confirmed by reported literature.
Step 3: 5-Phenyl-1-benzothiophene
##STR00256##
[0645] A solution of 5-bromo-1-benzothiophene (2 g, 9.385 mmol),
phenylboronic acid (1.2 g, 9.841 mmol), Pd(PPh.sub.3).sub.4, (544
g, 0.471 mmol), potassium carbonate (1.9 g, 13.747 mmol) in toluene
and water (20+20 mL) mixture was refluxed at 100.degree. C. for
overnight. After completion of the reaction, the mixture was cooled
to room temperature, diluted with ethyl acetate, filtered through a
Celite.RTM. bed, and washed with ethyl acetate. The combined
filtrate was washed with water and brine. The organic layer was
dried over sodium sulfate and concentrate under reduced pressure to
give residue which was purified by column chromatography using
hexane as eluent on silica gel to afford 5-phenyl-1-benzothiophene
(1.7 g, 8.084 mmol, 86%) as white solid. .sup.1H-NMR (400 MHz,
CDCl.sub.3): .delta. 7.95 (d, J=1.5 Hz, 1H), 7.86 (dd, J'=8.5 Hz,
J''=0.7 Hz, 1H), 7.60-7.58 (m, 2H), 7.52 (dd, J'=8.3 Hz, J''=1.7
Hz, 1H), 7.41-7.37 (m, 3H), 7.32-7.27 (m, 2H).
Step 4: 3-Bromo-5-phenyl-1-benzothiophene
##STR00257##
[0647] 5-Phenyl-1-benzothiophene (1.5 g, 7.132 mmol) was dissolved
in a mixture of chloroform (15 mL) and acetic acid (15 mL) and
cooled to 0.degree. C., and NBS (1.6 g, 8.989 mmol) was then added
portionwise. The reaction mixture was stirred at room temperature
for 48 hr. After completion of the reaction, the mass was quenched
by saturated solution of sodium thiosulfate and extracted with
chloroform. The combined organic layers were washed with saturated
solution of sodium bicarbonate and brine, dried over sodium sulfate
and concentrated under reduced pressure to give residue which was
purified by column chromatography using hexane as eluent on silica
gel to afford 3-bromo-5-phenyl-1-benzothiophene (1.9 g, 6.570 mmol,
92%) as yellow liquid. .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta.
8.04 (s, 1H), 7.90 (d, J=8.6 Hz, 1H), 7.70 (dd, J'=8.3 Hz, J''=1.5
Hz, 2H), 7.67 (dd, J'=8.3 Hz, J''=1.5 Hz, 1H), 7.50 (t, J=7.6 Hz,
2H), 7.48 (s, 1H), 7.42-7.39 (m, 1H).
Step 5: 3-Bromo-2-nitro-5-phenyl-1-benzothiophene
##STR00258##
[0649] A solution of 3-bromo-5-phenyl-1-benzothiophene (0.55 g,
1.901 mol) in acetic anhydride (2.28 mL) was cooled to 0.degree. C.
and a mixture of fuming nitric acid (0.54 mL) in acetic acid (0.37
mL) was added and the mass stirred for 2 hr. Progress of the
reaction was monitored by TLC. After completion of the reaction,
the mixture was quenched in ice-cold water, extracted with DCM,
dried over anhydrous sodium sulfate and concentrated under reduced
pressure to give residue which was purified by column
chromatography using hexane as eluent on silica gel to afford
3-bromo-2-nitro-5-phenyl-1-benzothiophene (0.15 g, 0.448 mmol, 34%)
as yellow solid.
[0650] .sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 8.18 (s, 1H),
7.86 (s, 2H), 7.66-7.63 (m, 2H), 7.51-7.47 (m, 2H), 7.43-7.39 (m,
1H).
Step 6:
N-(3-Chloro-4-fluorophenyl)-2-nitro-5-phenyl-1-benzothiophen-3-ami-
ne
##STR00259##
[0652] A solution of 3-bromo-2-nitro-5-phenyl-1-benzothiophene
(0.07 g, 0.209 mmol) and 3-chloro-4-fluoroaniline (0.061 g, 0.419
mmol) in DMF (1 mL) was heated to 120.degree. C. for 1 hr. After
completion of the reaction, the mixture was poured into ice-cold
water under stirring, solid precipitated, filtered, washed with
water, dried under vacuum and recrystallized with DCM and hexane
mixture to afford pure
N-(3-chloro-4-fluorophenyl)-2-nitro-5-phenyl-1-benzothiophen-3-amine
(0.05 g, 0.125 mmol, 60%) as orange solid. .sup.1H-NMR (400 MHz,
CDCl.sub.3): .delta. 10.03 (s, 1H), 7.74 (s, 2H), 7.46-7.44 (m,
1H), 7.38-7.35 (m, 2H), 7.33-7.31 (m, 2H), 7.28-7.25 (m, 2H),
7.24-7.22 (m, 2H).
Step 7:
N.sup.3-(3-Chloro-4-fluorophenyl)-5-phenyl-1-benzothiophene-2,3-di-
amine (Procedure E)
##STR00260##
[0654] To the solution of
N-(3-chloro-4-fluorophenyl)-2-nitro-5-phenyl-1-benzothiophen-3-amine
(0.05 g, 0.125 mmol) in methanol (3 mL) was added activated Pd/C
(0.005 g, 10%) under hydrogen gas (balloon) at room temperature and
the reaction mass stirred for 3 hr. The reaction mixture was
filtered through a Celite.RTM. bed under nitrogen, washed with
methanol, the filtrate was distilled to give residue which was
crystallized with a DCM and hexane mixture to afford
N.sup.3-(3-chloro-4-fluorophenyl)-5-phenyl-1-benzothiophene-2,3-diamine
(0.027 g, 0.073 mmol, 58%) as brown solid. .sup.1H-NMR (500 MHz,
DMSO-d.sub.6): .delta. 7.65 (d, J=8.3 Hz, 1H), 7.50 (d, J=7.4 Hz,
2H), 7.39-7.35 (m, 3H), 7.28-7.23 (m, 2H), 7.18 (s, 1H), 7.07 (t,
J=9.0 Hz, 1H), 6.54-6.52 (m, 1H), 6.47-6.44 (m, 1H), 5.94 (bs, 2H);
HRMS: 367.0471 [M-H].
Example 20
Synthesis of
2-Amino-3-((3-chloro-4-fluorophenyl)amino)benzo[b]thiophene-6-carbonitril-
e (Compound 136)
##STR00261##
[0655] Step 1: 1-Bromo-3-[(2,2-diethoxyethyl)sulfanyl]benzene
##STR00262##
[0657] To a stirred solution of 3-bromothiophenol (10.0 g, 52.88
mmol) in DMF (180 mL) was added NaH (3.0 g, 63.45 mmol) portionwise
at 0.degree. C. After the addition, the reaction mixture was
stirred at room temperature for 30 min. Bromoacetaldehyde
dimethylacetal (6.85 mL, 58.17 mmol) was added dropwise and the
resulting mixture was stirred at room temperature for 2 hr. The
reaction mixture was diluted with EtOAc (250 mL) and washed with
cold water and brine solution. The organic layer was dried over
anhydrous sodium sulfate, concentrated under reduced pressure to
afford the crude material which was purified by column
chromatography using silica gel (100-200 mesh) and 2% EtOAc-hexane
as eluent to afford
1-bromo-3-(2,2-dimethoxy-ethylsulphanyl)-benzene (11.0 g, 36.038
mmol, 68%) as colorless liquid. GCMS: 277 (m/z).
Step 2: 6-Bromobenzo[b]thiophene and 4-Bromobenzo[b]thiophene
##STR00263##
[0659] PPA (40.0 g) was combined with chlorobenzene (110 mL) and
refluxed for 1 hr. 1-Bromo-3-(2,2-dimethoxy-ethylsulphanyl)-benzene
(11.0 g, 36.038 mmol) in chlorobenzene (40 mL) was added dropwise
to the refluxing reaction mixture and continued to reflux for 12
hr. The reaction mixture was cooled to room temperature, the
top-layer was separated, the bottom layer was diluted with water
(200 mL) and then extracted with DCM (2.times.200 mL). The combined
organic layer was dried over anhydrous sodium sulfate, concentrated
under reduced pressure to afford the crude material which was
purified by column chromatography using silica gel (100-200 mesh)
and hexane as eluent to afford a mixture of
6-bromobenzo[b]thiophene and 4-bromo-benzo[b]thiophene (7.0 g,
32.849 mmol, 91%) as pale yellow liquid. GCMS: 213 (m/z).
Step 3: Benzo[b]thiophene-6-carbonitrile
##STR00264##
[0661] A mixture of 6-bromobenzo[b]thiophene and
4-bromobenzo[b]thiophene (5.0 g, 23.46 mmol) in DMF (90 mL) was
degassed with argon for 15 min. Pd(PPh.sub.3).sub.4 (1.35 g, 1.173
mmol) was then added and the mixture further degassed for 10 min.
Zn(CN)2 (2.75 g, 23.46 mmol) was then added and the mixture
refluxed for 3 hr. The mixture was allowed to cool to room
temperature and filtered through a pad of Celite.RTM. reagent. The
filtrate was concentrated, diluted with MTBE (150 mL), washed with
cold water, followed by brine. The organic layer was dried over
anhydrous sodium sulfate, concentrated under reduced pressure to
afford the crude material which was purified by column
chromatography using silica gel (100-200 mesh) and 2% EtOAc/hexane
as eluent to afford benzo[b]thiophene-6-carbonitrile (1.2 g, 7.537
mmol, 32%) as yellow liquid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 8.19 (s, 1H), 7.88 (d, 1H), 7.70 (d, 1H), 7.60-7.56 (m,
1H), 7.41 (d, 1H); GCMS: 159 (m/z).
Step 4: 3-Bromobenzo[b]thiophene-6-carbonitrile
##STR00265##
[0663] To a stirred solution of benzo[b]thiophene-6-carbonitrile
(1.0 g, 6.289 mmol) in CHCl.sub.3 (7 mL) and acetic acid (7 mL) was
added NBS (1.34 g, 7.547 mmol) portionwise at 0.degree. C. and the
mixture stirred at rt for 48 hr. The reaction mixture was diluted
with DCM (60 mL) and washed with saturated solution of sodium
thiosulfate solution and NaHCO.sub.3 followed by brine. The organic
layer was dried over sodium sulfate, concentrated under reduced
pressure to afford the crude material which was purified by column
chromatography using silica gel (100-200 mesh) and 2% EtOAc/hexane
as eluent to 3-bromobenzo[b]thiophene-6-carbonitrile (0.8 g, 3.359
mmol, 53%) as off white solid. GCMS: 238 (m/z).
Step 5: 3-Bromo-2-nitrobenzo[b]thiophene-6-carbonitrile
##STR00266##
[0665] To a stirred solution of
3-bromobenzo[b]thiophene-6-carbonitrile (0.5 g, 2.10 mmol) in DCM
(3 mL) was added trifluoroacetic anhydride (3.87 mL, 27.30 mmol) at
0.degree. C. and the mixture stirred for 30 min. KNO.sub.3 (0.23 g,
2.310 mmol) was then added portionwise at 0.degree. C. and the
mixture stirred for a further 30 min. The mixture was diluted with
DCM (25 mL) and washed with water and brine solution. The organic
layer was dried over sodium sulfate, concentrated under reduced
pressure to afford the crude material which was purified by
triturating with 50% MTBE and DCM to afford
3-bromo-2-nitro-benzo[b]thiophene-6-carbonitrile (0.25 g, 0.883
mmol, 42%) as yellow solid. LCMS: 281 (M-H).
Step 6:
3-((3-Chloro-4-fluorophenyl)amino)-2-nitrobenzo[b]thiophene-6-carb-
onitrile
##STR00267##
[0667] A solution of
3-bromo-2-nitrobenzo[b]thiophene-6-carbonitrile (0.8 g, 2.826 mmol)
and 4-fluoro-3-chloro phenyl amine (0.41 g, 2.826 mmol) in DMF (8
mL) was heated at 100.degree. C. in a sealed tube for 12 hr. The
mixture was diluted with MTBE (25 mL) and washed with cold water.
The organic layer was dried over sodium sulfate, concentrated under
reduced pressure to afford the crude material which was purified by
triturating with MTBE and pentane to afford
3-(3-chloro-4-fluoro-phenylamino)-2-nitrobenzo[b]thiophene-6-carbonitrile
(0.45 g, 1.294 mmol, 46%) as yellow solid. LCMS: 346.2 (M-H).
Step 7:
2-Amino-3-((3-chloro-4-fluorophenyl)amino)benzo[b]thiophene-6-carb-
onitrile (Procedure F)
##STR00268##
[0669] To a stirred solution of
3-(3-chloro-4-fluoro-phenylamino)-2-nitro-benzo[b]thiophene-6-carbonitril-
e (0.2 g, 0.575 mmol) in THF (2 mL) and methanol (2 mL) was added
zinc dust (0.075 g, 1.150 mmol) and NH.sub.4Cl (0.062 g, 0.575
mmol) at room temperature and the mixture stirred for 3 hr. The
mixture was filtered through a pad of Celite.RTM. reagent and
washed with ethyl acetate. The filtrate was diluted with EtOAc (30
mL) and washed with water and brine. The organic layer was dried
over sodium sulfate, concentrated under reduced pressure to afford
the crude material which was purified by triturating with MTBE and
pentane to afford 2-amino-3-(3-chloro-4-fluoro-phenylamino)-benzo
(b)thiophene-6-carbonitrile (0.08 g, 0.251 mmol, 44%) as pale brown
solid.
[0670] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 8.11 (s, 1H),
7.47 (d, J=8.3 Hz, 1H), 7.32 (s, 1H), 7.11 (t, J=9.1 Hz, 1H), 7.01
(d, J=8.2 Hz, 1H), 6.63 (bs, 2H), 6.51 (dd, J'=6.2 Hz, J''=2.6 Hz,
1H), 6.43-6.40 (m, 1H); LCMS: 316 (M-H).
Example 21
Synthesis of
(3-((3-Chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)-carbamate
(Compound 141) (Procedure G)
##STR00269##
[0672] To a stirred solution of
N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine
(1.0 eq) in THF was added pyridine (1.3 eq) followed by ethyl
chloroformate (1.1 eq) at 25.degree. C. The reaction mixture was
stirred for 3-10 hr at 25.degree. C. and concentrated. The crude
material was dissolved in EtOAc, washed thoroughly with water
followed by brine, dried over Na.sub.2SO.sub.4 and concentrated.
The crude mass was subjected to Prep-TLC/trituration to afford the
major desired mono-carbamate
(3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carbamate
(45-50%) along with traces of di-carbamate ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)furo[2,3-c]pyridin-3-y-
l)carbamate as solid.
Example 22
Synthesis of Ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)furo-[2,3-c]pyridin-3--
yl)carbamate (Compound 142) (Procedure H)
##STR00270##
[0674] To a stirred solution of
N.sup.3-(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine in
THF was added pyridine (10.0 eq) followed by ethyl chloroformate
(8.0 eq) at 25.degree. C. Reaction mixture was stirred for 20 hr at
25.degree. C. and concentrated, crude material was dissolved in
EtOAc, washed thoroughly with water followed by brine, dried over
Na.sub.2SO.sub.4 and concentrated. The crude mass was subjected to
Prep-TLC/trituration to afford the desired di-carbamate ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)furo[2,3-c]pyridin-3-y-
l)carbamate (50-55%) along with mono-carbamate
(343-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-2-yl)carbamate
(20-25%) as solid.
Example 23
Synthesis of
N.sup.3-(3-chloro-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine (Example 158)
##STR00271##
[0675] Step 1: 2-Bromo-3-(methoxymethoxy)pyridine
##STR00272##
[0677] To a stirred solution of 2-bromo-3-hydroxypyridine (50 g,
287.356 mmol) in THF at 0.degree. C. was added t-BuO-K (51.49 g,
459.7 mmol) portion wise. After stirring the reaction mixture for
15 mins, methoxymethyl chloride (34.473 mL, 459.77 mmol) was added
to it at 0.degree. C. and the resulting reaction mixture was
stirred for 12 h. at 25.degree. C. Reaction mixture was diluted
with water and extracted with ethyl acetate (4.times.500 mL).
Organic layer was dried over anhydrous sodium sulfate, concentrated
under reduced pressure to afford rude mass which was purified by
column chromatography using silica gel (100-200 mesh) and 10%
EtOAc-hexane as eluent to afford 2-bromo-3-methoxymethoxy-pyridine
(45 g) as pale brown liquid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.03 (dd, J'=4.5 Hz, J''=1.3 Hz, 1H), 7.60 (dd, J'=8.1 Hz,
J''=1.1 Hz, 1H), 7.40 (dd, J'=8.2 Hz, J''=4.5 Hz, 1H), 5.35 (s,
2H), 3.41 (s, 3H).
Step 2: 2-Bromo-3-(methoxymethoxy)isonicotinaldehyde
##STR00273##
[0679] To a stirred solution of 2-Bromo-3-Methoxymethoxypyridine
(10.0 g, 45.872 mmol) in anhydrous THF (140 mL) was added LDA (79.5
mL,59.633 mmol, 0.75 M in THF) at -78.degree. C. After stirring for
1 hr at -78.degree. C., ethylformate (5.559 mL, 68.807 mmol) was
added to it and stirred for 30 min at -78.degree. C. The cold bath
was removed and the reaction mixture was kept at -10.degree. C. and
quenched with aq. NH.sub.4Cl solution (50 mL). Reaction mass was
extracted with ethyl acetate (3.times.150 mL), dried over sodium
sulfate and was concentrated under reduced pressure to afford crude
mass which was passed through a small pad of silica gel (100-200
mesh) using 4% ethylacetate/hexane as eluent to get
2-bromo-3-methoxymethoxy-pyridine-4-carbaldehyde (5.0 g) as pale
yellow solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 10.2 (s,
1H), 8.40 (d, J=4.8 Hz, 1H), 7.67 (d, J=4.8 Hz, 1H), 5.25 (s, 2H),
3.55 (s, 3H)).
Step 3:
3-(Methoxymethoxy)-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde
##STR00274##
[0681] To a stirred solution of
2-bromo-3-methoxymethoxy-pyridine-4-carbaldehyde (8.0 g, 32.52
mmol), 2-methylpyridine-4-boronic acid (4.455 g, 32.52 mmol) and
tricyclohexyl phosphine (0.547 g, 1.951 mmol) in dry 1,4-dioxane
(500 mL) was added K.sub.3PO.sub.4 (55 mL, 1.27M), MeOH (50 mL) and
Pd.sub.2(dba).sub.3 (0.893 g, 0.976 mmol) under argon atmosphere.
The resulting reaction mixture was degassed with argon for 5 mins.
then reaction mixture was slowly heated to reflux for about 4 hrs.
After completion of reaction the reaction mass was cooled to room
temperature, filtered through celite bed, washed with ethyl
acetate, filtrate was concentrated under reduced pressure to give
crude mass which was purified by column chromatography on silica
gel (100-200 mess) using 60% EtOAc/hexane as eluent to afford
3-methoxymethoxy-2'-methyl-[2,4']bipyridinyl-4-carbaldehyde (6.0 g)
as brown liquid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 10.34
(s, 1H), 8.72 (d, J=4.7 Hz, 1H), 8.58 (d, J=5.1 Hz, 1H), 7.73 (d,
J=4.8 Hz, 1H), 7.69 (s, 1H), 7.63 (d, J=4.2 Hz, 1H), 4.96 (s, 2H),
3.20 (s, 3H), 2.56 (s, 3H).
Step 4: 3-Hydroxy-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde
##STR00275##
[0683] 3-Methoxymethoxy-2'-methyl-[2,4']bipyridinyl-4-carbaldehyde
(6.0 g, 65.83 mmol) was taken in DCM (10 mL) under ice bath and 10%
TFA-DCM (60 mL) was added slowly, resulting mixture was stirred at
25.degree. C. for 2 hrs. Reaction mixture was cooled under ice bath
and basified (pH 10) with potassium carbonate, extracted with ethyl
acetate (2.times.30 mL), aqueous part was neutralized with citric
acid and extracted with 10% IPA-DCM (5.times.50 mL). Combined
organic part was dried over sodium sulphate, concentrated under
reduced pressure to afford the crude mass which was purified by
trituration using mixture of solvents MTBE-pentane to give
3-hydroxy-2'-methyl-[2,4'-bipyridine]-4-carbaldehyde (4.0 g) as
brown solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 11.25
(bh, 1H), 10.29 (s, 1H), 8.55 (d, J=5.2 Hz, 1H), 8.42 (d, J=4.7 Hz,
1H), 7.86 (s, 1H), 7.81 (d, J=5.1 Hz, 1H), 7.69 (d, J=4.8 Hz, 1H),
2.55 (s, 3H); LCMS: 315.3 (M+H).
Step 5:
4-(3-Chloro-4-fluorophenyl)imino)methyl)-2'-methyl-[2,4'-bipyridin-
]-3-ol
##STR00276##
[0685] 3-Hydroxy-2'-methyl-[2,4']bipyridinyl-4-carbaldehyde (5.0 g,
23.364 mmol) was taken in mixed solvents of TFE (30 mL) and MeCN
(30 mL) and added 4-fluoro-3-chlorophenyl amine (3.4 g, 23.364
mmol) at 25.degree. C., resulting mixture was stirred at this
temperature for 2 hr. Reaction mixture was concentrated under
reduced pressure to give crude mass which was purified by
trituration with n-pentane to afford
4-{[3-chloro-4-fluoro-phenylimino]-methyl}-2'-methyl
[2,4']bipyridinyl-3-ol (6.0 g). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 9.22 (s, 1H), 8.78 (d, J=5.9 Hz, 1H), 8.52
(d, J=4.7 Hz, 1H), 8.39 (s, 1H), 8.32 (d, J=5.0 Hz, 1H), 7.95 (dd,
J=6.7 Hz, J''=2.3 Hz, 1H), 7.80 (d, .1=4.6 Hz, 1H), 7.65-7.57 (m,
2H), 2.71 (s, 3H); LCMS: 341.9 (M+H).
Step 6:
N.sup.3-(3-chloro-4-fluorophenyl)-7-(2-methylpyridin-4-y0furo[2,3--
c]pyridine-2,3-diamine
##STR00277##
[0687] To a stirred solution of
4-1[3-chloro-4-fluoro-phenylimino]-methyl1-2'-methyl-[2,4]bipyridinyl-3-o-
l (6.0 g, 17.595 mmol) in mixed solvent of DCM (36 mL) and TFE (36
mL) was added TMSCN (8.805 mL, 70.381 mmol) at 25.degree. C. The
reaction mixture was stirred for 12 h at 25.degree. C. After
completion of reaction the volatiles were removed and crude
material was purified by trituration with MTBE/pentane to afford
desired product
N.sup.3-(3-chloro-4-fluorophenyl)-7-(2-methylpyridin-4-yl)furo[2,3-c]pyri-
dine-2,3-diamine (3.0 g) as pale brown solid. .sup.1H-NMR (400 MHz,
CD.sub.3CN): .delta. 8.65 (s, 1H), 8.29-8.13 (m, 3H), 7.03 (bs,
2H), 6.66 (s, 1H), 6,60 (s, 1H), 5.94 (s, 1H), 5.78 (bs, 2H), 2.66
(s, 3H); LCMS: 369 (M+H).
Example 24
Synthesis of Ethyl
(7-((4-carbamoylphenyl)ethynyl)-2-((ethoxycarbonyl)
amino)furo[2,3-c]pyridin-3-yl)(3-chloro-4-fluorophenyl)carbamate
(Compound 160)
##STR00278##
[0688] Step 1: 3-Hydroxy-2-iodopyridine
##STR00279##
[0690] To a stirring solution of 3-hydroxy pyridine (30 g, 315.78
mmol) in water (3000 mL) was added Na.sub.2CO.sub.3 (70 g, 663.15
mmol) followed by Iodine (80 g, 315.78 mmol) in portion wise at
0.degree. C. The reaction mixture was allowed to stir at room
temperature for 2 hrs. Reaction mass was acidified with 1N HCl up
to pH -4 to get the solid which was filtered off, washed with
chilled water followed by MTBE-Hexane to afford desired
3-hydroxy-2-iodopyridine (45 g) as off white solid.
Step 2: 2-Iodo-3-(methoxymethoxy)pyridine
##STR00280##
[0692] To a stirred solution of 3-hydroxy-2-iodopyridine (10 g,
45.24 mmol) in THF: DMF (10 mL: 20 mL) at 0.degree. C. was added
tert-BuO-K (6 g, 54.3 mmol) portion wise. After stirring the
reaction mixture for 30 min, methoxymethyl chloride (4 mL, 50 mmol)
was added at 0.degree. C. and the resulting mixture was allowed to
stir at room temperature for 3 hrs. Reaction mixture was diluted
with brine and extracted with EtOAc (3.times.150 mL). EtOAc part
was dried over anhydrous sodium sulfate, concentrated under reduced
pressure to afford crude mass which was purified by column
chromatography using silica (100-200 mesh) and EtOAc-hexane as
eluent to afford 2-iodo-3-(methoxymethoxy)pyridine (6.0 g) as off
white solid.
Step 3: 2-Iodo-3-(methoxymethoxy)isonicotinaldehyde
##STR00281##
[0694] To a stirred solution of 2-iodo-3-(methoxymethoxy)pyridine
(4.0 g, 15.094 mmol) in anhydrous THF (60 mL) was added LDA (24.15
mL, 18.113 mmol, 0.75M in THF) drop wise at -78.degree. C. After
stirring the reaction mixture for 1 hr at -78.degree. C., DMF (1.74
mL, 22.642 mmol) was added to it and stirred for 15 mins at
-78.degree. C. Saturated ammonium chloride solution was added drop
wise into the reaction mass at -78.degree. C. and warm-up to room
temperature. The resulting quenched mass was extracted with ethyl
acetate (2.times.150 mL). EtOAc part was washed with water followed
by brine, dried over sodium sulfate and concentrated under reduced
pressure, to afford crude mass which was passed through a pad of
silica (100-200 mesh) using 3% EtOAc-hexane as eluent, to afford
2-iodo-3-(methoxymethoxy)-isonicotinaldehyde (2.2 g) as pale yellow
solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 10.14 (s, 1H),
8.39 (d, J=4.7 Hz, 1H), 7.60 (d, J=4.8 Hz, 1H), 5,22 (s, 2H), 3.55
(s, 3H).
Step 4: 3-Hydroxy-2-iodoisonicotinaldehyde
##STR00282##
[0696] To a stirred solution of
2-iodo-3-(methoxymethoxy)-isonicotinaldehyde (1.2 g, 4.096 mmol) in
DCM (3 mL) was added 10% TFA-DCM (10 mL) at 0.degree. C. and
stirred at room temperature for 3 hrs. Reaction mass was
concentrated under reduced pressure and crude was taken in water
(20 mL) and basified (pH-10) with potassium carbonate and extracted
with ethyl acetate (2.times.30 mL), aqueous part was neutralized
with citric acid and extracted with 10% IPA-DCM (5.times.50 mL).
Combined organic part was dried over sodium sulphate, concentrated
under reduced pressure to afford the crude material which was
purified by trituration using MTBE-pentane to afforded
3-hydroxy-2-iodoisonicotinaldehyde (0.75 g) as yellow solid.
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 11.45 (bh, 1H), 10.16
(s, 1H), 8.10 (d, J=3.7 Hz, 1H), 7.53 (d, J=3.4 Hz, 1H); LCMS:
249.5 (M+H).
Step 5:
N.sup.3-(3-Chloro-4-fluorophenyl)-7-iodofuro[2,3-c]pyridine-2,3-di-
amine
##STR00283##
[0698] To a stirred solution of 3-hydroxy-2-iodoisonicotinaldehyde
(4.5 g, 18.072 mmol) in dry DCM (70 mL) was added
3-chloro-4-fluoroaniline (2.63 g, 18.072 mmol), TMSCN (11.305 mL,
90.361 mmol) followed by TMSOTf (0.653 mL, 3.614 mmol) and the
reaction mass was stirred at room temperature for 5 hr. After
completion of the reaction, water was added to the reaction mass
and extracted with DCM (2.times.250 mL). The combined organic
layers were washed with water and saturated sodium bicarbonate
solution followed by brine, dried over sodium sulphate and
evaporated under reduced pressure to afford the crude mass which
was purified by trituration with MTBE/pentane to afford the desired
compound
N.sup.3-(3-chloro-4-fluorophenyl)-7-iodofuro[2,3-c]pyridine-2,3-diamine
(5.0 g) as brown solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 7.83 (d, J=5.0 Hz, 1H), 7.21 (s, 2H), 7.15-7.09 (m, 2H),
6.83 (d, J=5.0 Hz, 1H), 6.57-6.56 (m, 1H), 6.47-6.45 (m, 1H); LCMS:
404.0 (M+H).
Step 6: Ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-iodofuro
[2,3-c]pyridin-3-yl)carbamate
##STR00284##
[0700] To a stirred solution of
N.sup.3-(3-chloro-4-fluorophenyl)-7-iodofuro[2,3-c]pyridine-2,3-diamine
(5.0 g, 12.407 mmol) in dry THF (50 mL), pyridine (8.011 mL, 99.256
mmol) was added and the reaction mass was stirred for 15 min, then
ethylchloroformate (11.811 mL, 124.069 mmol) was added to it.
Reaction mass was allowed to stir at room temperature for 4 hrs.
After completion of the reaction, the reaction mass was diluted
with water and extracted with EtOAc (2.times.100 mL). The organic
layer was washed with water followed by brine, dried over sodium
sulphate and evaporated under reduced pressure to afford the crude
material which was purified by trituration with MTBE/pentane to
afford the desired compound ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-iodofuro[2,3-c]pyri-
din-3-yl)carbamate (95.5 g) as brown solid. .sup.1H-NMR (400 MHz,
CD.sub.3CN): .delta. 8.20 (d, J=5.2 Hz, 1H), 7.32 (d, J=5.1 Hz,
1H), 7.10 (t, J=9.0 Hz, 1H), 6.84 (dd, J'=6.2 Hz, J''=2.8 Hz, 1H),
6.75 (dt, J'=8.8 Hz, J''=6.8 Hz, J''=3.1 Hz, 1H), 6.56 (s, 1H),
4.21 (q, J=7.1 Hz, 4H), 1.17 (t, J=7.1 Hz, 6H); LCMS: 548
(M+H).
Step 7: Ethyl
(7-((4-carbamoylphenyl)ethynyl)-2-((ethoxycarbonyl)amino)
furo[2,3-c]pyridine-3-yl)(3-chloro-4-fluorophenyl)carbamate
##STR00285##
[0702] To a stirred solution of ethyl
(3-chloro-4-fluorophenyl)(2-((ethoxycarbonyl)amino)-7-iodofuro[2,3-c]pyri-
din-3-yl)carbamate (0.2 g, 0.366 mmol) in triethylamine (5 mL) and
THF (2 mL) was added 4-ethynylbenzamide (0.058 g, 0.402 mmol), CuI
(0.003 g, 0.018 mmol), the resulting mass was degassed for 20 mins
then PdCl.sub.2(PPh.sub.3).sub.2 (0.013 g, 0.018 mmol) was added
and again degassed for another 10 mins. Reaction mass was heated to
50.degree. C. for 3 hrs. After completion of reaction the reaction
mass was cooled to room temperature and filtered through celite
bed. Filtrate was concentrated under reduced pressure to give
residue which was diluted with water and extracted with ethyl
acetate (3.times.50 mL), EtOAc part was washed with brine, dried
over sodium sulfate and concentrated under reduced pressure to
afford crude product which was purified by column chromatography on
silica (100-200 mesh) using ethyl acetate/hexane as an eluent to
afford desired product ethyl
(7-((4-carbamoylphenypethynyl)-2-((ethoxycarbonyl)amino)furo[2,3-c]pyridi-
ne-3-yl)(3-chloro-4-fluorophenyl)carbamate (60 mg) as brown solid.
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 8.47 (d, J=5.1 Hz,
1H), 8.36 (s, 1H), 8.12 (s, 1H), 7.97 (d, J=8.0 Hz, 2H), 7.76 (d,
J=8.0 Hz, 2H), 7.52 (s, 1H), 7.49 (d, J=5.1 Hz, 1H), 7.26 (t, J=9.0
Hz, 1H), 6.82 (dd, J'=6.2 Hz, J''=2.5 Hz, 1H), 6.75-6.71 (m, 1H),
4.18 (q, J=7.0 Hz, 4H), 1.11 (t, J=7.0 Hz, 6H); LCMS: 563.2
(M-H).
Example 25
Synthesis of
N'-(3-Chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]pyridine-
-2,3-diamine (Compound 180)
##STR00286##
[0703] Step 1: 6-Fluoro-3-hydroxy-2-iodopyridine
##STR00287##
[0705] To a stirring solution of 6-fluoro-3-hydroxypyridine (10 g,
88.496 mmol) in THF (300 mL) was added slowly Na.sub.2CO.sub.3
(19.697 g, 185.841 mmol) in water (300 mL) at 0.degree. C. and
stirred for 20 mins. Iodine (22.461 g, 88.496 mmol) was added
portion wise at 0.degree. C. and the reaction mass was allowed to
stir at room temperature for overnight. Solvent was evaporated;
aqueous part was neutralized with 1N HCl, extracted with ethyl
acetate. Organic layer was separated, dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under reduce pressure to give crude
mass which was triturated with DCM/pentane to get
6-fluoro-3-hydroxy-2-iodopyridine (19.8 g) as a brown solid.
.sup.1H-NMR (400 MHz, DMSO): .delta. 10.79 (s, 1H), 7.31-7.27 (m,
1H), 7.03-7.00 (m, 1H); LCMS: 239.8 (M+H).
Step 2: 6-Fluoro-2-iodo-3-(methoxymethoxy)pyridine
##STR00288##
[0707] To a stirred solution of 6-fluoro-3-hydroxy-2-iodopyridine
(9.0 g, 37.657 mmol) in THF (180 mL) at 0.degree. C. was added
tert-BuO-K (6.76 g, 60.251 mmol) portion-wise. After stirring the
reaction mixture for 30 min, methoxy methyl chloride (4.55 mL,
60.251 mmol) was added at 0.degree. C. and the resulting mixture
was stirred for 2 hr at room temperature. After completion of
reaction the reaction mass was diluted with brine and extracted
with EtOAc (3.times.150 mL). The combined organic layers were
washed with brine, dried over anhydrous sodium sulfate,
concentrated under reduced pressure to afford crude mass (9.6 g)
which was purified by column chromatography using silica (100-200
mesh) and 5% EtOAc-Hexane as eluent to afford
6-fluoro-2-iodo-3-(methoxymethoxy)pyridine (8.0 g) as yellow solid.
LCMS: 284 (M+H).
Step 3: 6-Fluoro-3-(methoxymethoxy)-2,4'-binyridine
##STR00289##
[0709] To a stirred solution of
6-fluoro-2-iodo-3-(methoxymethoxy)pyridine (3.5 g, 12.367 mmol) in
dioxane (120 mL) was added 4-pyridinylboronic acid (1.67 g, 13.6
mmol), K.sub.3PO.sub.4 (16.5 mL, 21.02 mmol, 1.27 M solution in
water), P(Cy).sub.3 (0.7 g, 2.47 mmol). The reaction mass was
degassed for 20 min. with Argon then Pd.sub.2(dba).sub.3 (1.13 g,
1.24 mmol) was added and degassed for another 10 min. The resulting
mixture was heated to 100.degree. C. for 2 hrs. Reaction mass was
cooled to room temperature and filtered through celite bed,
filtrate was concentrated under reduced pressure to give crude mass
which was purified by column chromatography to afford
6-fluoro-3-(methoxymethoxy)-2,4'-bipyridine (2.8 g) as pale yellow
solid. .sup.1H-NMR (400 MHz, DMSO): .delta. 8.69 (d, J=5.0 Hz, 2H),
7.94-7.89 (m, 3H), 7.28 (dd, J'=3.7 Hz, J''=8.9 Hz, 1H), 5.33 (s,
2H), 3.34 (s, 3H); LCMS: 235.2 (M+H).
Step 4:
6-Fluoro-3-(methoxymethoxy)-[2,4'-bipyridine]-4-carbaldehyde
##STR00290##
[0711] To a stirred solution of
6-fluoro-3-(methoxymethoxy)-2,4'-bipyridine (3.0 g, 12.8 mmol) in
anhydrous THF (40.0 mL) was added n-BuLi (7 mL, 14.1 mmol, 2.0 M in
hexane) drop wise at -78.degree. C. After stirring for 1 hr at
-78.degree. C., DMF (1.5 mL, 19.2 mmol) was added to it and stirred
for 15 mins at -78.degree. C. Saturated ammonium chloride solution
was added drop wise to the reaction mass at -78 C and warm-up to
room temperature. Reaction mass was extracted with ethyl acetate
(2.times.150 mL), The combined organic layers were washed with
water followed by brine, dried over sodium sulfate and concentrated
under reduced pressure to afford crude mass
6-fluoro-3-(methoxymethoxy)-[2,4'-bipyridine]-4-carbaldehyde (3.5
.mu.m) which was used as such in the next step.
Step 5: 6-Fluoro-3-hydroxy-[2,4'-bipyridine]-4-carbaldehyde
##STR00291##
[0713] To a stirred solution of crude
6-fluoro-3-(methoxymethoxy)-[2,4'-bipyridine]-4-carbaldehyde (3.5
g, 13.35 mmol) in DCM (5.0 mL) was added 10% TFA-DCM solution (50.0
mL) at 0.degree. C. Reaction mass was allowed to stir at room
temperature for 5 hrs. Evaporated the volatiles, residue was
diluted with water (20 mL) and basified (pH-10) with potassium
carbonate and extracted with ethyl acetate (2.times.30 mL), aqueous
part was neutralized with citric acid and extracted with 10%
IPA-DCM (5.times.150 mL), combined organic part was dried over
sodium sulphate, concentrated under reduced pressure to give crude
mass which was purified by trituration using MTBE-pentane to afford
6-fluoro-3-hydroxy-[2,4'-bipyridine]-4-carbaldehyde (1.5 g) as
yellow solid. .sup.1H-NMR (400 MHz, DMSO): .delta. 11.15 (bh, 1H),
10.27 (s, 1H), 8.72 (d, J=5.3 Hz, 2H), 8.01 (d, J=4.9 Hz, 2H), 7.49
(d, J=3.6 Hz, 2H).
Step 6:
N.sup.3-(3-Chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,-
3-c]pyridine-2,3-diamine
##STR00292##
[0715] To a stirred solution of
6-fluoro-3-hydroxy-[2,4'-bipyridine]-4-carbaldehyde (0.5 g, 0.23
mmol) in DCM (7 mL) was added 4-fluoro-3-chloroaniline (0.333 g,
0.2.3 mmol), TMS-CN (1.5 mL, 12 mmol) and TMS-OTf (0.08 mL, 0.45
mmol) at 25.degree. C. The resulting reaction mixture was stirred
at this temperature for 20 hr. Reaction mass was concentrated and
diluted with ethyl acetate (25 mL) and washed with water
(2.times.25 mL) followed by brine solution and dried over sodium
sulphate, concentrated under reduce pressure to give crude mass
which was taken in DCM (2 mL), and added TMSCN (0.5 mL), TMSOTf
(0.02 mL) and the resulting mixture was kept in the refrigerator
for 16 hrs. Volatiles were removed and diluted with ethyl acetate
(25 mL), washed with water (2.times.25 mL) followed by brine
solution and dried over sodium sulphate, concentrated and purified
by trituration using ethyl acetate/DCM to afforded
N.sup.3-(3-Chloro-4-fluorophenyl)-5-fluoro-7-(pyridin-4-yl)furo[2,3-c]pyr-
idine-2,3-diamine (210 mg) as pale yellow solid. .sup.1H-NMR (400
MHz, DMSO-d.sub.6): .delta. 8.76 (d, J=5.8 Hz, 2H), 8.15 (d, J=5.8
Hz, 2H), 7.50 (s, 2H), 7.17-7.12 (m, 2H), 6.63 (dd, J=6.1 Hz,
J''=2.5 Hz, 1H), 6.53-6.49 (m, 2H); LCMS: 373.1 (M+H).
Example 26
Synthesis of
N.sup.3-(3-Chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine-2,3-dia-
mine (Example 190)
##STR00293##
[0716] Step 1:
2,6-Difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
##STR00294##
[0718] 2,6 Difluoropyridine (5.0 g, 43.44 mmol) was taken in dry
THF (60 mL) under Nitrogen atmosphere and cooled to -78.degree. C.
then n-BuLi (26.06 mL, 52.1739 mmol 2.0 M in hexane) was added
drop-wise over 30 min., followed by B(O.sup.iPr).sub.3 (11.095 mL,
47.785 mmol) at the same temperature and maintained 20 min.
Resulting reaction mixture was allowed to warm to room temperature
and added pinacol (5.647 g, 47.785 mmol) and acetic acid (4.969 mL,
86.881 mmol). The resulting reaction mixture was stirred at room
temperature for 3 h. After completion of reaction, saturated
aqueous NH.sub.4Cl (100 mL) was poured into reaction mass and
extracted with ethyl acetate (3.times.150 mL). Combined organic
layer was washed with saturated aq. NaHCO.sub.3 solution (50 mL)
followed by brine solution (2.times.50 mL), dried over anhydrous
Na.sub.2SO.sub.4 and concentrated to afford crude material which
was passed through a pad of silica to afford
2,6-difluoro-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
(9.0 g) as a brown gummy solid and use as such in next step.
Step 2: 2,6-Difluoro-3-hydroxypyridine
##STR00295##
[0720] To a stirred solution of sodium perborate tetrahydrate (17.2
g, 112.03 mmol) in water (130 mL) was added THF (130 mL) and 2,6-di
fluoro-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine
(9.0 g, 37.344 mmol) at 0.degree. C. Resulting reaction mixture was
stirred at room temperature for 3 h. Reaction mixture was extracted
with ethyl acetate (3.times.150 mL), washed with brine (150 mL) and
dried over sodium sulfate and concentrated under reduced pressure
to afford crude material which was purified by column
chromatography on silica gel (100-200 mesh) using 30% EtOAc-hexane
as eluent to afford 2,6-difluoro-3-hydroxypyridine (4.0 g) as white
solid. LCMS: 130 (M-H).
Step 3: 2,6-Difluoro-3-(methoxymethoxy)pyridine
##STR00296##
[0722] To a stirred solution of 2,6-difluoro-3-hydroxypyridine (4.0
g, 30.534 mmol) in DCM (80 mL) at 0.degree. C. was added DIPEA
(10.6 mL,61.069 mmol), after stirring the reaction mixture for 15
min, methoxy methyl chloride (3.46 mL, 45.802 mmol) was added to it
at 0.degree. C. and the resulting mixture was stirred for 3 h at
room temperature. Water was added to the reaction mixture and
extracted with DCM (2.times.50 mL). The combined organic part was
washed with brine (150 mL) and dried over sodium sulfate and
concentrated under reduced pressure to afford crude mass which was
purified by column chromatography on silica gel (100-200 mesh
silica) using 5% EtOAc-hexane as eluent to afford
2,6-difluoro-3-methoxymethoxy-pyridine (3.2 g) as brown liquid.
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 7.94 (m, 1H), 7.13
(dd, J'=8.5 Hz, J''=2.8 Hz, 1H), 5.27 (s, 2H), 3.41 (s, 3H).
Step 3: 2,6-Difluoro-3-(methoxymethoxy)isonicotinaldehyde
##STR00297##
[0724] To a stirred solution of
2,6-difluoro-3-methoxymethoxy-pyridine (3.5 g, 20.0 mmol) in
anhydrous THF (60 mL) was cooled to -78.degree. C. then added
n-BuLi (11 mL, 22 mmol, 2.0 M in hexane) drop-wise maintaining the
temperature -78.degree. C. After stirring for 1 h at -78.degree.
C., DMF (2.3 mL, 30 mmol) was added and stirred for 30 min at
-78.degree. C. Reaction mixture was warm to -10.degree. C. and
quenched by adding saturated solution of NH.sub.4Cl (30 mL)
drop-wise. Reaction mass was extracted with ethyl acetate
(3.times.50 mL), washed with brine, dried over sodium sulfate and
concentrated under reduced pressure to afford
2,6-difluoro-3-methoxymethoxy-pyridine-4-carbaldehyde (4.0 g) as a
brown liquid which was used as such in the next step.
Step 4: 2,6-Difluoro-3-hydroxyisonicotinaldehyde
##STR00298##
[0726] To a stirred solution of
2,6-difluoro-3-methoxymethoxy-pyridine-4-carbaldehyde (4.0 g,
19.704 mmol) in DCM (10 mL) was added 10% TFA-DCM solution (100 mL)
at 0.degree. C. and stirred at room temperature for 5 h. Reaction
mixture was concentrated, diluted with water and neutralized by
solid K.sub.2CO.sub.3 (pH 7), extracted with 5% IPA-DCM (3.times.50
mL). Combined organic layer was washed with water, followed by
brine solution, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated under reduce pressure to afford the crude material
which was purified by column chromatography on (100-200 mesh
silica) using 30% EtOAc-hexane as eluent to afford
2,6-difluoro-3-hydroxyisonicotinaldehyde (1.5 g) as brown liquid.
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 11.45 (bh, 1H), 10.31
(d, J=2.6 Hz, 1H), 7.19 (dd, J'=8.0 Hz, J''=2.6 Hz, 1H).
Step 4:
4-(((3-Chloro-4-fluorophenyl)imino)methyl)-2,6-difluoropyridin-3-o-
l
##STR00299##
[0728] To a stirred solution of
2,6-difluoro-3-hydroxyisonicotinaldehyde (1.0 g, 6.289 mmol) in a
mixed solvent [TFE(5 mL):MeCN(5 mL)] was added
4-fluoro-3-chloroaniline (0.915 g, 6.289 mmol) at room temperature,
resulting reaction mixture was stirred at room temperature for 2 h.
After completion of reaction, the reaction mass was concentrated
and triturated with n-pentane to afford
4-(((3-chloro-4-fluorophenyl)imino)methyl)-2,6-difluoropyridin-3-ol
(1.3 g) as a yellow solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 11.93 (s, 1H), 9.00 (s, 1H), 7.79 (d, J=6.4 Hz, 1H),
7.57-7.53 (M, 2H), 7.39 (s, 1H).
Step 4:
N.sup.3-(3-Chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine--
2,3-diamine
##STR00300##
[0730] To a stirred solution of
4-(((3-chloro-4-fluorophenyl)imino)methyl)-2,6-difluoropyridin-3-ol
(1.3 g, 4.545 mmol) in a mixed solvent [TFE (5 mL):DCM (5 mL)] was
added TMSCN (2.95 mL, 23.636 mmol) followed by TMSOTf (0.164 mL,
0.909 mmol) at room temperature and allowed to stir at room
temperature for 16 h. Reaction mass was concentrated and residue
was taken in DCM, washed with NaHCO.sub.3 solution, followed by
brine solution, dried over anhydrous sodium sulfate and
concentrated under reduced pressure to give crude material was
purified by column chromatography on silica gel (100-200 mesh) 30%
EtOAc-hexane as eluent to
affordN.sup.3-(3-chloro-4-fluorophenyl)-5,7-difluorofuro[2,3-c]pyridine-2-
,3-diamine (0.4 g) as brown solid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 7.46 (s, 2H), 7.16-7.11 (m, 2H), 6.60 (dd,
J'=6.2 Hz, J''=2.7 Hz, 1H), 6.48 (dt, J'=8.8 Hz, J''=6.8 Hz,
J''=3.4 Hz, 1H), 6.42 (s, 1H); LCMS: 314 (M+H).
Example 27
Synthesis of
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7-yl)-N-m-
ethylpicolinamide (Example 195)
##STR00301##
[0731] Step 1: 4-Chloro-N-methylpicolinamide
##STR00302##
[0733] To 4-chloro-pyridine-2-carboxylic acid (5.0 g, 31.847 mmol)
was added SOCl.sub.2 (15 mL) and refluxed for 3 h. The volatiles
were removed and the reaction mixture was diluted with THF (220
mL), added Et.sub.3N (13.42 mL, 95.541 mmol) and MeNH.sub.2 (19.1
mL, 2M in THF) at 0.degree. C. The reaction mixture was stirred at
room temperature for 12 h. After completion of reaction, volatiles
were removed to give residue which was diluted with ethyl acetate,
washed with saturated solution of sodium bicarbonate. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4 and evaporated
under reduce pressure to afford 4-chloro-pyridine-2-carboxylic acid
methylamide (4.6 g) as brown liquid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.83 (bs, 1H), 8.62 (d, J=5.2 Hz, 1H), 8.01
(d, J=2.0 Hz, 1H), 7.75 (dd, J'=5.3 Hz, J''=2.0 Hz, 1H), 2.82 (d,
J=4.8 Hz, 3H); LCMS: 171 (M+H).
Step 2:
N-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinami-
de
##STR00303##
[0735] A degassed mixture of 4-chloropyridine-2-carboxylic acid
methylamide (4.6 g, 27.059 mmol), bis(pinacolato)diboron (8.246 g,
32.471 mmol), Pd(OAC).sub.2 (0.243 g, 1.082 mmol), PCy.sub.3 (0.379
g, 1.353 mmol) and potassium acetate (3.983 g, 40.588 mmol) in
1,4-dioxane (215 mL) was heated at 100.degree. C. under nitrogen
for 4 h. The reaction mixture was filtered through celite bed and
the filtrate was concentrated in vacuum to get crude mass
N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
(6.2 g) as pale brown liquid which was used in next step as such
without further purification. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.75 (d, J=4.4 Hz, 1H), 8.61 (d, J=4.5 Hz, 1H), 8.20 (s,
1H), 7.71 (d, J=4.4 Hz, 1H), 2.82 (d, J=4.8 Hz, 3H), 1.28 (s,
12H).
Step 3:
4-Formyl-3-(methoxymethoxy)-N-methyl-[2,4'-bipyridine]-2'-carboxam-
ide
##STR00304##
[0737] To a stirred solution of
2-bromo-3-methoxymethoxy-pyridine-4-carbaldehyde (5.0 g, 20.325
mmol) in 1,4-dioxane (250 mL) was added crude
N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
(3.659 g, 20.325 mmol), K.sub.3PO.sub.4 (27.2 mL, 34.553 mmol, 1.27
M in water) and P(Cy).sub.3 (1.14 g, 4.065 mmol). The reaction
mixture was degassed for 20 min with Argon then added
Pd.sub.2(dba).sub.3 (1.86 g, 2.033 mmol) and again degassed for
another 5 min. The reaction mixture was heated to 100.degree. C.
for 2 h. After completion of reaction the reaction mixture was cool
to room temperature, the volatiles were removed under reduced
pressure to afford crude
4-formyl-3-(methoxymethoxy)-N-methyl-[2,4'-bipyridine]-2'-carboxamide
(6.3 g), which was forwarded to the next step as such. LCMS: 302
(M+H).
Step 4:
4-Formyl-3-hydroxy-N-methyl-[2,4'-bipyridine]-2'-carboxamide
##STR00305##
[0739] 10% TFA-DCM (60 mL) solution was added to crude
4-formyl-3-(methoxymethoxy)-N-methyl-[2,4'-bipyridine]-2'-carboxamide
(6.1 g, 20.266 mmol) in DCM (6 mL) at 0.degree. C. After stirring
the reaction mixture for 3 h at room temperature, concentrated
under reduced pressure, diluted with water and was basified using
solid potassium carbonate, washed with ethyl acetate and the
aqueous part was acidified to pH-6 using citric acid and extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous sodium sulfate and concentrated under reduce
pressure to afford crude mass which was purified by trituration
using DCM/Et2O/pentane gave pure
4-formyl-3-hydroxy-N-methyl-[2,4'-bipyridine]-2'-carboxamide (2.8
g) as pale brown solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 11.26 (s, 1H), 10.31 (s, 1H), 8.84 (d, J=4.6 Hz, 1H), 8.75
(d, J=5.0 Hz, 1H), 8.67 (s, 1H), 8.51 (d, J=4.7 Hz, 1H), 8.17 (dd,
J'=5.0 Hz, J''=1.6 Hz, 1H), 7.76 (d, J=4.8 Hz, 1H), 2.85 (d, J=4.8
Hz, 3H); LCMS: 258.2 (M+H).
Step 5:
4-(2-Amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-7--
yl)-N-methylpicolinamide
##STR00306##
[0741] To a stirred solution of
4-formyl-3-hydroxy-N-methyl-[2,4'-bipyridine]-2'-carboxamide (0.5
g, 1.946 mmol) in DCM (6.0 mL) was added 3-chloro-4-fluoroaniline
(0.283 g, 1.946 mmol), TMSCN (1.268 mL, 10.117 mmol), TMSOTf (0.071
mL, 0.389 mmol) at room temperature in a sealed tube. The reaction
mixture was stirred for 1 hr at 40.degree. C., followed by addition
of 10 mmol NH.sub.4OAc buffer (3.0 mL) and stirred for 12 h. The
reaction mixture was filtered through a sintered funnel and washed
the solid with MTBE/hexane/10-20%EA-hexane to remove trace
impurities to get desired compound,
4-(2-amino-3-((3-chloro-4-fluorophenyl)amino)furo[2,3-c]pyridin-
-7-yl)-N-methylpicolinamide (0.450 g) as orange solid. .sup.1H-NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.88 (s, 1H), 8.83-8.78 (m, 2H),
8.40 (d, J=4.4 Hz, 1H), 8.25 (d, J=4.9 Hz, 1H), 7.18-7.11 (m, 4H),
6.98 (d, J=4.9 Hz, 1H), 6.62-6.60 (m, 1H), 6.53-6.51 (m, 1H), 2.88
(d, J=4.6 Hz, 3H); LCMS: 412 (M+H).
Example 28
Synthesis of
N.sup.3-(3-Chloro-4-fluorophenyl)-N.sup.7,N.sup.7-diphenylfuro[2,3-e]pyri-
dine-2,3,7-triamine (Example 200)
##STR00307##
[0742] Step 1: 3-(Methoxymethoxy)-N,N-diphenylpyridin-2-amine
##STR00308##
[0744] A solution of 2-bromo-3-methoxymethoxy-pyridine (2.0 g,
9.174 mmol), diphenylamine (2.018 g, 11.927 mmol), ter-BuONa (1.763
g, 18.349 mmol) and Dppf (0.305 g, 0.55 mmol) in toluene (27.5 mL)
was degassed for 20 min. and in this degassed reaction mixture was
added Pd.sub.2 (dba).sub.3 (0.168 g, 0.183 mmol). The resulting
reaction mixture was again degassed for 10 min. and heated at
80.degree. C. for 16 h. After completion of reaction the reaction
mass was filtered through celite bed and the filtrate was
evaporated under reduce pressure to obtained crude mass which was
purified by column chromatography on silica gel (100-200 mesh)
using 20% EtOAc/hexane as eluent to afford
(3-methoxymethoxy-pyridin-2-yl)-diphenyl-amine (2.0 g) as brown
solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 7.99 (dd, J=4.7
Hz, J''=1.4 Hz, 1H), 7.53 (dd, J'=4.7 Hz, J''=1.4 Hz, 1H),8.1 Hz,
J''=1.4 Hz, 1H), 7.26-7.19 (m, 5H), 6.98 (t, J=7.3 Hz, 2H), 6.85
(d, J=7.6 Hz, 4H), 5.01 (s, 2H), 3.01 (s, 3H).
Step 2: 2-(Diphenylamino)-3-(methoxymethoxy)isonicotinaldehyde
##STR00309##
[0746] To a stirred solution of
(3-methoxymethoxy-pyridin-2-yl)-diphenyl-amine (1.0 g, 3.264 mmol)
in THF (13 mL) was added n-BuLi (1.8 mL, 3.6 mmol, 2 Min hexane)
drop wise at -78 .degree. C. After stirring the reaction mixture
for 1 h at -78.degree. C., DMF (0.377 mL, 4.896 mmol) was added and
stirred for 15 min at -78.degree. C. Reaction mass was quenched
with saturated solution of NH.sub.4Cl and extracted with ethyl
acetate (2.times.100 mL). Combined organic layer was washed with
brine, dried over anhydrous sodium sulfate, concentrated under
reduced pressure to afford crude
2-diphenylamino-3-methoxymethoxy-pyridine-4-carbaldehyde (1.0 g)
which was forwarded to the next step without further purification.
LCMS: 335 (M+H).
Step 3: 2-(Diphenylamino)-3-hydroxyisonicotinaldehyde
##STR00310##
[0748] To a stirred solution of crude
2-diphenylamino-3-methoxymethoxy-pyridine-4-carbaldehyde (1.0 g,
2.991 mmol) in DCM (5 mL) was added 10% TFA-DCM (15 mL) at
0.degree. C. After stirring the reaction mixture for l h at room
temperature, the volatiles were evaporated under reduced pressure.
Thus obtained crude was diluted with water (15 mL) and basified up
to pH-10 by potassium carbonate and washed with MTBE. The aqueous
part was neutralized with citric acid and extracted with 10%
IPA-DCM (2.times.100 mL). The organic layer was separated, dried
over anhydrous sodium sulfate, concentrated under reduced pressure
to afford 2-diphenylamino-3-hydroxy-pyridine-4-carbaldehyde (0.5 g)
as orange solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 10.56
(s, 1H), 10.27 (s, 1H), 8.02 (d, J=4.8 Hz, 1H), 7.42 (d, J=4.9 Hz,
1H), 7.26 (t, J=7.8 Hz, 4H), 7.02 (t, J=7.2 Hz, 2H), 6.90 (d, J=7.9
Hz, 4H); LCMS: 291 (M+H).
Step 4:
4-(((3-Chloro-4-fluorophenyl)imino)methyl)-2-(diphenylamino)pyridi-
n-3-ol
##STR00311##
[0750] To a solution of
2-diphenylamino-3-hydroxy-pyridine-4-carbaldehyde (0.5 g, 1.722
mmol) in a mixed solvent of triflouroethanol (4 mL) and
acetonitrile (4 mL) was added 3-chloro-4-flouroaniline (0.25 g,
1.722 mmol) and the mixture was stirred for 5 h at room
temperature. Reaction mass was evaporated to dryness under reduced
pressure to afford
4-(((3-chloro-4-fluorophenyl)imino)methyl)-2-(diphenylamino)pyridin-3-ol
(0.7 g) as reddish brown liquid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 12.74 (s, 1H), 9.06 (s, 1H), 8.02 (d, J=4.9
Hz, 1H), 7.86-7.84 (m, 1H), 7.53-7.49 (m, 2H), 7.47 (d, J=5.0 Hz,
1H), 7.26 (t, J=7.7 Hz, 4H), 7.00 (t, J=7.3 Hz, 2H), 6.89 (d, J=7.9
Hz, 4H).
Step 5:
N.sup.3-(3-Chloro-4-fluorophenyl)-N.sup.7,N.sup.7-diphenylfuro[2,3-
-c]pyridine-2,3,7-triamine
##STR00312##
[0752] To a stirred solution of
4-(((3-chloro-4-fluorophenyl)imino)methyl)-2-(diphenylamino)pyridin-3-ol
(0.7 g,1.675 mmol) in a mixed solvent of trifluoroethanol (5 mL)
and dichloromethane (5 mL) was added TMSCN (1.09 mL, 8.711 mmol) at
room temperature and stirred for 16 h at ambient temperature. After
completion of reaction the solvent was evaporated and thus obtained
crude material was purified by column chromatography followed by
triturating with ether/pentane to afforded
N.sup.3-(3-chloro-4-fluorophenyl)-N.sup.7,N.sup.7-diphenylfuro[2,3-c]pyri-
dine-2,3,7-triamine (0.15 g) as brown solid. .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 7.82 (d, J=5.1 Hz, 1H), 7.27 (t, J=7.8 Hz,
4H), 7.14 (t, J=9.0 Hz, 1H), 7.05-7.01 (m, 3H), 6.93 (d, J=7.8 Hz,
4H), 6.72 (d, J=5.0 Hz, 1H), 6.67 (bs, 2H), 6.59 (dd, J'=6.3 Hz,
J''=2.5 Hz, 1H), 6.49-6.44 (m, 1H); LCMS: 445 (M+H).
Example 29
Synthesis of
N.sup.3-(3-Chlorophenyl)-7-(2-methylpyridin-4-yl)benzo[b]thiophene-2,3-di-
amine (Compound 248)
##STR00313##
[0753] Step 1: (2-Bromophenyl)(2,2-diethoxyethyl)sulfane
##STR00314##
[0755] To a stirred suspension of K.sub.2CO.sub.3 (9.56 g, 69.17
mmol) in acetone (40 mL) was added 2-bromothiophenol (10.0 g, 53.21
mmol) at 0.degree. C. and stirred the reaction mixture at same
temperature for 20-30 min. then added bromoacetaldehyde
diethylacetal (11.5 g, 58.33 mmol) drop-wise and stirred the
reaction mass at 25.degree. C. for 18 h. The reaction was monitor
by tic and after completion of reaction volatiles were remove under
reduce pressure to get concentrated mass which was diluted with
water, extracted with ethyl acetate (3.times.150 mL), combined
organic layers were washed with water then brine, dried over
anhydrous sodium sulfate, concentrated under reduced pressure to
give crude mass which was purified by column chromatography on
silica gel using 5% ethyl acetate/hexane mixture as eluent to
afford (2-bromophenyl)(2,2-diethoxyethyl)sulfane (15.6 g) as pale
yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 7.49 (d,
J=7.8 Hz, 1H), 7.32 (d, J=7.8 Hz, 1H), 7.21 (t, J=7.6 Hz, 1H), 6.98
(t, J=7.6 Hz, 1H), 4.67 (t, J=5.5 Hz, 1H), 3.65 (q, J=6.9 Hz, 2H),
3.52 (q, J=6.8 Hz, 2H), 3.12 (d, J=6.0 Hz, 2H), 1.17 (t, J=6.9 Hz,
6H).
Step 2: 7-Bromobenzo[b]thiophene
##STR00315##
[0757] To a stirred solution of PPA (42.22 g) in chlorobenzene (100
mL) was added solution of (2-bromophenyl)(2,2-diethoxyethyl)sulfane
(26.2 g, 86.185 mmol) in chlorobenzene (20 mL) drop-wise at
130.degree. C. and allowed the reaction mass to stirred for 4 h.
After completion of reaction the reaction mixture was cool down to
room temperature and decants the organic layer and keeps it aside.
The residue was extracted with toluene (3.times.100 mL) and
combined organic layers were evaporated under reduced pressure to
give crude mass which was purified by column chromatography on
silica gel using 2% ethyl acetate/hexane as eluent to afford
7-bromobenzo[b]thiophene (12.7 g, 59.62 mmol) as colorless liquid.
.sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 7.76 (dd, J'=8.2 Hz,
J''=0.9 Hz, 1H), 7.50-7.47 (m, 2H), 7.42 (d, J=5.5 Hz, 1H), 7.23
(t, J=7.8 Hz, 1H).
Step 3: 3,7-Dibromobenzo[b]thiophene
##STR00316##
[0759] N-Bromosuccinimide (5.73 g. 32.2 mmol) was added slowly into
the stirred solution of 7-bromobenzo[b]thiophene (5.28 g, 24.78
mmol) in chloroform (25 mL) and acetic acid (25 mL) at 0.degree. C.
The resulting reaction mixture was stirred at 25.degree. C. for 24
h. The reaction was monitor by tic and after completion of reaction
the reaction mixture washed with saturated solution of
Na.sub.2S.sub.2O.sub.3 (20 mL), NaHCO.sub.3 (20 mL) and water (10
mL). The organic part was dried over anhydrous sodium sulfate and
concentrated under reduce pressure to give crude product which was
purified by column chromatography on silica gel using hexane as
eluent to afford 3,7-dibromobenzo[b]thiophene (4.7 g, 16.135 mmol)
as white solid. .sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 7.80
(dd, J'=8.2 Hz, J''=0.9 Hz, 1H), 7.57 (d, J=7.8 Hz, 1H), 7.52 (s,
1H), 7.36 (t, J=7.8 Hz, 1H).
Step 4: 3,7-Dibromo-2-nitrobenzo[b]thiophene
##STR00317##
[0761] To a stirred solution of 3,7-dibromobenzo[b]thiophene (4.7
g, 16.09 mmol) in acetic anhydride (30 mL) was added fuming
HNO.sub.3 followed by acetic acid (3.2 g, 53.9 mmol) dropwise at
5-10.degree. C. Reaction mixture was stirred at same temperature
for 2 h then quenched with ice-water, the solid was
precipitate-out, filtered and washed with water, dried under vacuum
to give crude material which was purified by crystallization using
DCM/hexane to afford 3,7-dibromo-2-nitrobenzo[b]thiophene (1.63 g,
4.839 mmol), as yellow solid. .sup.1H-NMR (400 MHz, CDCl.sub.3):
.delta. 8.02 (d, J=8.2 Hz, 1H), 7.79 (d, J=7.8 Hz, 1H), 7.49 (t,
J=8.0 Hz, 1H).
Step 4:
7-Bromo-N-(3-chlorophenyl)-2-nitrobenzo[b]thiophen-3-amine
##STR00318##
[0763] To a stirred solution of
3,7-dibromo-2-nitrobenzo[b]thiophene (0.6 g, 1.78 mmol), in DMF (5
mL) was added 3-chloroaniline (0.681 g, 5.34 mmol) at room
temperature. The reaction mixture was stirred at 80.degree. C. for
2 h. After completion of reaction the reaction mass was quenched
with ice-water, the solid was precipitate, filtered and washed with
water to give wet solid which was dried under hot air oven to
afford desired product
7-bromo-N-(3-chlorophenyl)-2-nitrobenzo[b]thiophen-3-amine (0.250
g, 0.652 mmol) as yellow solid. .sup.1H-NMR (400 MHz, CDCl.sub.3):
.delta. 9.88 (s, 1H), 7.65 (d, J=7.8 Hz, 1H), 7.38-7.29 (m, 3H),
7.18-7.15 (m, 2H), 7.04 (t, J=8.0 Hz, 1H); HRMS: 380.910 (M-H).
Step 5:
N-(3-Chlorophenyl)-7-(2-methylpyridin-4-yl)-2-nitrobenzo[b]thiophe-
n-3-amine
##STR00319##
[0765] To a stirred solution of
7-bromo-N-(3-chlorophenyl)-2-nitrobenzo[b]thiophen-3-amine (0.25 g,
0.651 mmol) and 2-methyl-4-pyridinylboronic acid (0.116 g, 0.847
mmol) in DMF (5 mL) and H.sub.2O (1 mL) mixture was added
K.sub.3PO.sub.4 (0.415 g, 1.955 mmol) and Pd(PPh.sub.3).sub.4
(0.075 g, 0.065 mmol, 10 mol %) at room temperature. The resulting
reaction mixture was stirred at 100.degree. C. for 2 h, completion
of reaction was monitor by tic. After completion of reaction the
reaction mass was cooled to room temperature, filtered through
celite bed and filtrate was extracted with ethyl acetate
(3.times.20 mL), washed with brine, dried over sodium sulfate,
concentrated under reduced pressure to give crude mass which was
purified by column chromatography on silica gel using 15%
EtOAc/hexane as eluent to afford
N-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)-2-nitrobenzo[b]thio-
phen-3-amine (0.07 g, 0.176 mmol) as reddish solid. .sup.1H-NMR
(400 MHz, CDCl.sub.3): .delta. 9.93 (s, 1H), 8.67 (d, J=5.3 Hz,
1H), 7.56-7.51 (m, 2H), 7.48 (d, J=5.0 Hz, 1H), 7.40-7.34 (m, 2H),
7.32-7.28 (m, 3H), 7.19 (d, J=7.6 Hz, 1H), 2.75 (s, 3H); HRMS:
396.057 (M+H).
Step 6:
N.sup.3-(3-Chlorophenyl)-7-(2-methylpyridin-4-yl)benzo[b]thiophene-
-2,3-diamine
##STR00320##
[0767] To a stirred solution of
N-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)-2-nitrobenzorbithiophen-3-ami-
ne (0.06 g, 0.152 mmol) in methanol (5 mL) was added activated Pd/C
(0.02 g, 10% Pd on charcoal) under nitrogen gas at room temperature
then nitrogen gas was replaced by hydrogen gas balloon and continue
stirring at ambient temperature for 2 h. After completion of
reaction the reaction mixture was filtered through celite bed
washed with methanol and filtrate was evaporated under reduce
pressure to give crude mass which was purified by trituration using
pentane to afford
N.sup.3-(3-chlorophenyl)-7-(2-methylpyridin-4-yl)benzo[b]thiophene-2,3-di-
amine (0.02 g, 0.054 mmol, 36%) as off white solid. .sup.1H-NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.53 (d, J=5.3 Hz, 1H), 7.52 (s,
1H), 7.45 (d, J=4.8 Hz, 1H), 7.40 (s, 1H), 7.25 (t, J=7.7 Hz, 1H),
7.10-7.06 (m, 1H), 7.04-6.99 (m, 2H), 6.55 (d, J=7.8 Hz, 1H), 6.45
(d, J=8.0 Hz, 1H), 6.42 (s, 1H), 5.98 (s, 2H), 2.52 (s, 3H); HRMS:
366.08 (M+H).
[0768] Compounds of the invention made according to procedures A-K
and Examples 1-29 as described herein are listed below in TABLE
1.
TABLE-US-00004 TABLE 1 .sup.1H-NMR (400 No. Structure IUPAC Name
MHz) proton shift LCMS Proc. 1 ##STR00321## N.sup.3-(3-Chloro-
4-fluorophenyl) furo[3,2- b]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.06 (d, J = 4.8 Hz, 1H), 7.52 (d, J = 7.8 Hz, 1H),
7.12-7.07 (m, 2H), 6.85 (dd, J' = 7.7 Hz, J'' = 5.2 Hz, 1H), 6.77
(bs, 2H), 6.56-6.55 (m, 1H), 6.51-6.49 (m, 1H) [M + H] 278.0 B 2
##STR00322## N.sup.3-(3-Chloro- 4-fluorophenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06
(d, J = 5.1 Hz, 1H), 7.15- 7.10 (m, 2H), 6.91 (s, 2H), 6.86 (d, J =
5.1 Hz, 1H), 6.56- 6.54 (m, 1H), 6.48- 6.45 (m, 1H) [M - H] 276.2 A
3 ##STR00323## N.sup.3- Phenylfuro [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.03 (d, J = 5.1 Hz, 1H), 7.07
(t, J = 7.7 Hz, 2H), 6.84-6.79 (m, 4H), 6.59 (t, J = 7.3 Hz, 1H),
6.52 (d, J = 7.9 Hz, 2H) [M - H] 224.0 A 4 ##STR00324## N.sup.3-(2-
Chlorophenyl) furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.42 (s, 1H), 8.04 (d, J = 5.0 Hz, 1H), 7.30 (d, J = 7.7
Hz, 1H), 7.02 (t, J = 7.7 Hz, 1H), 6.92 (bs, 2H), 6.81 (d, J = 5.0
Hz, 1H), 6.65- 6.62 (m, 2H), 6.31 (d, J = 8.1 Hz, 1H). [M + H]
260.0 A 5 ##STR00325## N.sup.3-(4- Fluorophenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.03
(d, J = 4.9 Hz, 1H), 6.91 (t, J = 8.6 Hz, 2H), 6.84- 6.82 (m, 4H),
6.51- 6.50 (m, 2H) [M - H] 242.0 A 6 ##STR00326## N.sup.3-(4-
(Trifluoro- methyl)phenyl) furo[2,3-c] pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 8.43 (s, 1H), 8.05 (d, J = 5.0 Hz, 1H), 7.56
(s, 1H), 7.40 (d, J = 8.3 Hz, 2H), 7.03 (bs, 2H), 6.87 (d, J = 5.0
Hz, 1H), 6.63 (d, J = 8.2 Hz, 2H) [M - H] 292.0 B 7 ##STR00327##
N.sup.3-(2,4- Dichloro- phenyl)furo [2,3-c]pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 8.45 (s, 1H), 8.06 (s, 1H), 7.42 (s, 1H),
7.11 (bs, 3H), 6.85 (s, 2H), 6.30 (d, J = 8.9 Hz, 1H) [M + H] 294.0
A 8 ##STR00328## N.sup.3-(3- Chlorophenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06
(d, J = 5.0 Hz, 1H), 7.20 (s, 1H), 7.09 (t, J = 8.2 Hz, 1H), 6.91
(s, 2H), 6.85 (d, J = 5.0 Hz, 1H), 6.61 (d, J = 7.6 Hz, 1H), 6.48
(m, 2H) [M - H] 258.2 A 9 ##STR00329## N.sup.3-(4-Chloro-3-
methoxyphenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.42 (s, 1H), 8.06 (d, J = 4.6 Hz, 1H), 7.10 (s, 1H), 7.05
(d, J = 8.6 Hz, 1H), 6.95 (bs, 2H), 6.86 (d, J = 5.0 Hz, 1H), 6.34
(d, J = 1.7 Hz, 1H), 6.04 (dd, J' = 8.4 Hz, J'' = 2.0 Hz, 1H), 3.68
(s, 3H) [M + H] 290.0 A 10 ##STR00330## N.sup.3-(2-Bromo-4-
fluorophenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.42 (s, 1H), 8.05 (d, J = 4.2 Hz, 1H), 7.44 (dd, J' = 8.2
Hz, J'' = 2.9 Hz, 1H), 6.99-6.95 (m, 3H), 6.80 (d, J = 5.0 Hz, 1H),
6.37 (s, 1H), 6.28 (dd, J' = 9.1 Hz, J'' = 5.3 Hz, 1H) [M - H]
320.0 A 11 ##STR00331## N.sup.3-(4- Morpholino- phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s, 1H), 8.02 (d,
J = 4.0 Hz, 1H), 6.81-6.72 (m, 5H), 6.47 (bs, 3H), 3.69 (bs, 4H),
2.89 (bs, 4H) [M + H] 311.0 A 12 ##STR00332## N.sup.3-(2,4,5-
Trifluoro- phenyl)furo [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.42 (s, 1H), 8.07 (d, J = 4.9 Hz, 1H), 7.47-7.40 (m, 1H),
7.04 (s, 1H), 6.95 (s, 2H), 6.89 (d, J = 5 Hz, 1H), 6.24- 6.17 (m,
1H) [M + H] 280.0 A 13 ##STR00333## N.sup.3-(3,4- Dimethyl-
phenyl)furo [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.38 (s, 1H), 8.02 (d, J = 5 Hz, 1H), 6.83-6.79 (m, 2H), 6.75 (s,
2H), 6.56 (s, 1H), 6.35 (d, J = 2.1 Hz, 1H), 6.24 (dd, J' = 8.1 Hz,
J'' = 2.3 Hz, 1H), 2.06 (s, 6H) [M - H] 252.0 A 14 ##STR00334##
N.sup.3-(3,5- Dimethyl- phenyl)furo [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.04 (d, J = 5.1 Hz, 1H), 6.83
(d, J = 5.1 Hz, 1H), 6.77 (bs, 2H), 6.67 (s, 1H), 6.24 (s, 1H),
6.14 (s, 2H), 2.10 (s, 6H) [M + H] 254.0 A 15 ##STR00335##
N.sup.3-(3,4- Dichloro- phenyl)furo [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06 (d, J = 4.8 Hz, 1H), 7.34
(s, 1H), 7.28 (d, J = 8.8 Hz, 1H), 6.97 (bs, 2H), 6.87 (d, J = 4.9
Hz, 1H), 6.64 (s, 1H), 6.49 (d, J = 8.2 Hz, 1H) [M - H] 291.8 A 16
##STR00336## N.sup.3-(4- Bromophenyl) furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.04 (d, J = 5.0 Hz,
1H), 7.21 (d, J = 8.5 Hz, 2H), 7.08 (s, 1H), 6.88 (bs, 2H), 6.82
(d, J = 5.0 Hz, 1H), 6.47 (d, J = 8.5 Hz, 2H) [M + H] 304.0 A 17
##STR00337## N.sup.3-(4- Chlorophenyl) furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.04 (d, J = 5.0 Hz,
1H), 7.10 (d, J = 8.4 Hz, 2H), 7.05 (s, 1H), 6.86 (bs, 2H), 6.82
(d, J = 4.8 Hz, 1H), 6.52 (d, J = 8.4 Hz, 2H) [M + H] 260.0 A 18
##STR00338## N.sup.3-(3-Chloro-4- iodophenyl)furo [2,3-c]pyridine-
2,3-diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06 (d, J = 5.0
Hz, 1H), 7.54 (d, J = 8.6 Hz, 1H), 7.35 (s, 1H), 6.95 (s, 2H), 6.86
(d, J = 4.9 Hz, 1H), 6.67 (bs, 1H), 6.29 (d, J = 6.9 Hz, 1H) [M -
H] 383.8 A 19 ##STR00339## N.sup.3-(3- Ethylphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.04 (d,
J = 4.6 Hz, 1H), 6.97 (t, 1H), 6.83-6.77 (m, 4H), 6.46-6.40 (m,
2H), 6.38 (m, 1H), 2.43 (merged in DMSO peak, 2H), 1.10 (t, J = 7.3
Hz, 3H) [M + H] 254.0 A 20 ##STR00340## N.sup.3-(3- Iodophenyl)
furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.40 (s,
1H), 8.06 (d, J = 5.0 Hz, 1H), 7.10 (s, 1H), 6.93-6.84 (m, 6H),
6.50 (d, J = 7.8 Hz, 1H) [M + H] 351.9 A 21 ##STR00341##
N.sup.3-(3-Bromo-4- methylphenyl) furo[2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.04 (d, J = 5.0 Hz, 1H), 7.03
(d, J = 8.2 Hz, 1H), 6.97 (s, 1H), 6.88 (bs, 2H), 6.83 (d, J = 5.0
Hz, 1H), 6.69 (m, 1H), 6.46-6.43 (m, 1H), 2.18 (s, 3H) [M - H]
316.0 A 22 ##STR00342## N.sup.3-(3-Bromo- 4,5- difluorophenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s,
1H), 8.07 (d, J = 5.0 Hz, 1H), 7.35 (s, 1H), 6.98 (bs, 2H), 6.89
(d, J = 4.9 Hz, 1H), 6.52 (s, 1H), 6.45-6.40 (m, 1H) [M + H] 340.0
A 23 ##STR00343## N.sup.3-(4- Benzylphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s, 1H), 8.02 (d,
J = 5.1 Hz, 1H), 7.27-7.23 (m, 2H), 7.18-7.12 (m, 3H), 6.93 (d, J =
8.2 Hz, 2H), 6.81 (d, J = 5.0 Hz, 1H), 6.75 (bs, 2H), 6.72 (s, 1H),
6.45 (d, J = 8.2 [M + H] 316.2 A Hz, 2H), 3.76 (s, 2H) 24
##STR00344## N.sup.3-(2- (Piperidin-1- yl)phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.02 (d,
J = 5.1 Hz, 1H), 7.03 (d, J = 7.5 Hz, 1H), 6.84- 6.77 (m, 3H), 6.74
(d, J = 5.1 Hz, 1H), 6.61 (t, J = 7.2 Hz, 1H), 6.20 (d, J = 7.8 Hz,
1H), 5.99 (s, 1H), 2.87 (bs, 4H), 1.73-1.69 (m, 4H), 1.54 (bs, 2H)
[M - H] 307.0 A 25 ##STR00345## N.sup.3-(5-Chloro-2- methoxyphenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.06 (d, J = 5.1 Hz, 1H), 6.89-6.87 (m, 3H), 6.82 (d, J = 5.1
Hz, 1H), 6.60 (dd, J' = 8.4 Hz, J'' = 2.5 Hz, 1H), 6.55 (s, 1H),
3.06 (d, J = 3.5 Hz, 1H), 3.85 (s, 3H) [M + H] 290.4 A 26
##STR00346## N.sup.3-(3- (Trifluoro- methyl)phenyl) furo[2,3-c]
pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.42 (s, 1H), 8.06 (d,
J = 5.0 Hz, 1H), 7.36 (s, 1H), 7.29 (t, J = 7.9 Hz, 1H), 6.93-6.89
(m, 3H), 6.86 (d, J = 5.1 Hz, 1H), 6.80 (s, 1H), 6.73 (d, J = 8.3
Hz, 1H) [M + H] 294.2 A 27 ##STR00347## N.sup.3-(4-Fluoro-3-
methylphenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.39 (s, 1H), 8.04 (d, J = 5.0 Hz, 1H), 6.86-6.81 (m, 4H),
6.71 (s, 1H), 6.38-6.36 (m, 1H), 6.32-6.29 (m, 1H), 2.09 (s, 3H) [M
+ H] 257.8 A 28 ##STR00348## 3-((2- Aminofuro [2,3-c]pyridin-
3-yl)amino)-2- fluoro- benzonitrile DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.06 (d, J = 4.8 Hz, 1H), 7.38 (s, 1H), 7.09-6.99 (m, 4H),
6.89 (d, J = 4.6 Hz, 1H), 6.62 (t, J = 7.6 Hz, 1H) [M + H] 269.0 A
29 ##STR00349## N.sup.3-(4- Phenoxyphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s, 1H), 8.05 (d,
J = 5.0 Hz, 1H), 7.29 (t, J = 7.7 Hz, 2H), 6.99 (t, J = 7.2 Hz,
1H), 6.87-6.82 (m, 8H), 6.55 (d, J = 8.5 Hz, 2H) [M - H] 316.1 A 30
##STR00350## N.sup.3-(2-Chloro-4- fluorophenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.05 (d,
J = 5.0 Hz, 1H), 7.30 (dd, J' = 8.5 Hz, J'' = 2.8 Hz, 1H),
6.95-6.90 (m, 3H), 6.81 (d, J = 5.1 Hz, 1H), 6.57 (s, 1H), 6.28
(dd, J' = 9.0 Hz, J'' = 5.3 Hz, 1H) [M + H] 278.0 A 31 ##STR00351##
N.sup.3-(2,3,4- Trichloro- phenyl)furo [2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.42 (s, 1H), 8.06 (d, J = 5.0 Hz,
1H), 7.28 (d, J = 9.0 Hz, 1H), 7.11 (s, 1H), 7.01 (bs, 2H), 6.86
(d, J = 5.2 Hz, 1H), 6.29 (d, J = 9.0 Hz, 1H). [M + H] 327.8 A 32
##STR00352## N.sup.3-(3,5-Di-tert- butylphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.03 (bs,
1H), 6.84 (bs, 1H), 6.72- 6.67 (m, 4H), 6.40 (s, 2H), 1.33-1.18 (m,
18H) [M + H] 338.2 A 33 ##STR00353## N.sup.3-(3-Chloro-4-
methylphenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.40 (s, 1H), 8.05 (d, J = 5.0 Hz, 1H), 7.03 (d, J = 8.2
Hz, 1H), 6.97 (s, 1H), 6.86 (bs, 2H), 6.83 (d, J = 5.0 Hz, 1H),
6.50 (s, 1H), 6.42 (d, J = 8.1 Hz, 1H), 2.12 (s, 3H) [M + H] 274.0
A 34 ##STR00354## N.sup.3-(2,4- Dimethoxy- phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s, 1H), 8.01 (d,
J = 5.0 Hz, 1H), 6.75 (d, J = 5.1 Hz, 1H), 6.72 (bs, 2H), 6.56 (d,
J = 2.4 Hz, 1H), 6.26 (dd, J' = 8.6 Hz, J'' = 2.5 Hz, 1H), 6.07 (d,
J = 8.5 Hz, 1H), 5.78 (s, 1H), 3.84 (s, 3H), 3.64 (s, 3H) [M + H]
286.0 A 35 ##STR00355## N.sup.3- (Benzo[d][1,3] dioxol-5-
yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s,
1H), 8.03 (d, J = 4.9 Hz, 1H), 6.83 (d, J = 5.0 Hz, 1H), 6.79 (bs,
2H), 6.65-6.63 (m, 2H), 6.17 (s, 1H), 5.95 (d, J = 8.1 Hz, 1H),
5.83 (s, 2H) [M + H] 270.0 A 36 ##STR00356## N.sup.3-(4-
Ethynylphenyl) furo[2,3-c] pyridine-2,3- diamine CD.sub.3CN:
.delta. 8.43 (s, 1H), 8.12 (d, J = 5.1 Hz, 1H), 7.25 (d, J = 8.6
Hz, 2H), 6.95 (d, J = 4.9 Hz, 1H), 6.56 (d, J = 8.6 Hz, 2H), 5.87
(s, 1H), 5.36 (s, 2H), 3.18 (s, 1H) [M + H] 250.0 A 37 ##STR00357##
N.sup.3-(3- Isopropoxy- phenyl)furo [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.04 (d, J = 5.0 Hz, 1H), 6.95
(t, J = 8.2 Hz, 1H), 6.84- 6.80 (m, 4H), 6.14 (dd, J' = 15.2 Hz,
J'' = 7.9 Hz, 2H), 6.00 (s, 1H), 4.43- 4.37 (m, 1H), 1.18 [M + H]
284.2 A (d, J = 6 Hz, 6H) 38 ##STR00358## N.sup.3-(3,5- Dimethoxy-
phenyl)furo [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.39 (s, 1H), 8.05 (d, J = 4.8 Hz, 1H), 6.89 (s, 1H), 6.84 (d, J =
5.0 Hz, 1H), 6.79 (s, 2H), 5.80 (s, 1H), 5.71 (s, 2H), 3.60 (s, 6H)
[M - H] 284.0 A 39 ##STR00359## 4-((2- Aminofuro[2,3- c]pyridin-3-
yl)amino) benzonitrile DMSO-d.sub.6: .delta. 8.42 (s, 1H), 8.06 (d,
J = 5.1 Hz, 1H), 7.84 (s, 1H), 7.49 (d, J = 8.7 Hz, 2H), 7.01 (s,
2H), 6.84 (d, J = 5.0 Hz, 1H), 6.60 (d, J = 8.3 Hz, 2H) [M + H]
251.0 A 40 ##STR00360## N.sup.3-(p- tolyl)Furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.02 (d, J = 5.1 Hz,
1H), 8.88 (d, J = 8.2 Hz, 2H), 6.78 (d, J = 5.0 Hz, 1H), 6.76 (s,
2H), 6.65 (s, 1H), 6.43 (d, J = 8.2 Hz, 2H), 2.15 (s, 3H) [M + H]
240.0 A 41 ##STR00361## 1-(4-((2- Aminofuro[2,3- c]pyridin-3-
yl)amino) phenyl) ethanone DMSO-d.sub.6: .delta. 8.42 (s, 1H), 8.06
(d, J = 5.0 Hz, 1H), 7.75-7.71 (m, 3H), 6.96 (bs, 2H), 6.83 (d, J =
5.0 Hz, 1H), 6.56 (d, J = 8.5 Hz, 2H), 2.40 (s, 3H) [M + H] 268.2 A
42 ##STR00362## N.sup.3-(3- Cyclopropyl- phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.03 (d,
J = 5.0 Hz, 1H), 6.93 (t, J = 7.6 Hz, 1H), 6.81 (d, J = 5.1 Hz,
1H), 6.76-6.74 (m, 3H), 6.31-6.26 (m, 3H), 1.74-1.71 (m, 1H),
0.85-0.81 (m, 2H), [M + H] 266.2 A 0.53-0.49 (m, 2H) 43
##STR00363## N.sup.3-(3- Isopropyl- phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H), 8.03 (d,
J = 5.1 Hz, 1H), 6.97 (t, J = 7.7 Hz, 1H), 6.82 (d, J = 5.1 Hz,
1H), 6.77 (bs, 3H), 6.49-6.46 (m, 2H), 6.28 (d, J = 7.6 Hz, 1H),
2.74- 2.67 (m, 1H), 1.12 [M + H] 268.2 A (d, J = 6.9 Hz, 6H) 44
##STR00364## N.sup.3-(3-Chloro- 2,4- difluorophenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06
(d, J = 4.9 Hz, 1H), 7.03-6.88 (m, 5H), 6.29-6.23 (m, 1H) [M + H]
296.0 A 45 ##STR00365## N.sup.3-(2,3- Dichloro-4- fluorophenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.05 (s, 1H), 7.13 (bs, 1H), 7.00 (s, 2H), 6.87 (s, 2H), 6.27
(s, 1H) [M + H] 312.0 A 46 ##STR00366## N.sup.3-(3- Fluorophenyl)
furo[2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.05 (d, J = 5.0 Hz,
1H), 7.19 (s, 1H), 7.09 (q, J = 7.7 Hz, 1H), 6.88 (s, 2H), 6.85 (d,
J = 5.0 Hz, 1H), 6.38-6.35 (m, 2H), 6.22 (d, J = 12.2 Hz, 1H) [M +
H] 244.0 A 47 ##STR00367## N.sup.3-(3,5- Dichloro-4- fluorophenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.08 (d, J = 5.0 Hz, 1H), 7.36 (s, 1H), 7.01 (bs, 2H), 6.90
(d, J = 5.0 Hz, 1H), 6.55 (d, J = 5.6 Hz, 2H) [M + H] 312.0 A 48
##STR00368## N.sup.3-(m- tolyl)Furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6): .delta. 8.40 (s, 1H), 8.04 (d, J = 5.1 Hz, 1H), 6.95
(t, J = 7.7 Hz, 1H), 6.82 (d, J = 5.1 Hz, 1H), 6.80 (bs, 2H), 6.75
(s, 1H), 6.41 (d, J = 7.2 Hz, 1H), 6.35- 6.31 (m, 2H), 2.14 [M + H]
240.0 A (s, 3H) 49 ##STR00369## N.sup.3-(Pyridin-3- yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.05
(s, 1H), 7.97 (s, 1H), 7.82 (s, 1H), 7.13 (s, 1H), 7.07 (bs, 1H),
6.93 (bs, 2H), 6.85 (d, J = 3.6 Hz, 1H), 6.74 (d, J = 7.2 Hz, 1H)
[M + H] 227.0 A 50 ##STR00370## N.sup.3-(4- (Trifluoro- methoxy)
phenyl)furo [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.40 (s, 1H), 8.05 (d, J = 4.7 Hz, 1H), 7.13 (s, 1H), 7.07 (d, J =
8.1 Hz, 2H), 6.88 (bs, 2H), 6.85 (d, J = 4.7 Hz, 1H), 6.55 (d, J =
8.5 Hz, 2H) [M + H] 310.2 A 51 ##STR00371## N.sup.3-(5-Chloro- 2,4-
difluorophenyl) furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.42 (s, 1H), 8.07 (d, J = 5.0 Hz, 1H), 7.46-7.41 (m, 1H),
7.06 (s, 1H), 6.97 (bs, 2H), 6.90 (d, J = 5.0 Hz, 1H), 6.29 (t, J =
8.1 Hz, 1H) [M + H] 296.0 A 52 ##STR00372## N.sup.3-(5-
Chloropyridin- 3-yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.43 (s, 1H), 8.08 (d, J = 5.1 Hz, 1H), 7.92 (s, 1H), 7.83
(s, 1H), 7.53 (s, 1H), 7.04 (bs, 2H), 6.91 (d, J = 4.9 Hz, 1H),
6.73 (s, 1H) [M - H] 259.0 A 53 ##STR00373## N.sup.3-([1,1'-
Biphenyl]-3- yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.39 (s, 1H), 8.02 (d, J = 5.1 Hz, 1H), 7.62 (d, J = 7.5
Hz, 2H), 7.48 (t, J = 7.6 Hz, 2H), 7.38 (t, J = 7.5 Hz, 1H),
7.08-7.03 (m, 2H), 6.84-6.82 (m, 3H), 6.73 (t, J = 7.3 Hz, [M + H]
302.2 A 1H), 6.34 (d, J = 8.0 Hz, 1H), 5.80 (s, 1H) 54 ##STR00374##
N.sup.3-(2,4- Dimethyl- phenyl)furo [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.02 (d, J = 5.1 Hz, 1H), 6.84
(s, 1H), 6.78 (d, J = 5.1 Hz, 1H), 6.74 (bs, 2H), 6.68 (d, J = 8.4
Hz, 1H), 6.09 (d, J = 8.1 Hz, 1H), 5.96 (s, 1H), 2.23 (s, 3H), 2.13
(s, 3H) [M + H] 254.0 A 55 ##STR00375## N.sup.3-(2,5- Dichloro-
phenyl)furo [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.43 (s, 1H), 8.07 (d, J = 5.1 Hz, 1H), 7.33 (d, J = 8.4 Hz, 1H),
7.02 (bs, 2H), 6.99 (s, 1H), 6.86 (d, J = 5.1 Hz, 1H), 6.68 (dd, J'
= 8.4 Hz, J'' = 2.4 Hz, 1H), 6.22 (d, J = 2.4 Hz, 1H) [M + H] 294.0
A 56 ##STR00376## (2-Amino-3- ((3-chloro-4- fluorophenyl) amino)-7-
methylfuro[2,3- c]pyridin-4- yl)methanol DMSO-d.sub.6: .delta. 7.94
(s, 1H), 7.12 (t, J = 9.0 Hz, 1H), 6.89 (s, 1H), 6.78 (bs, 2H),
6.54 (d, J = 3.7 Hz, 1H), 6.45-6.43 (m, 1H), 4.92 (s, 1H), 4.45 (s,
2H), 2.50 (s, 3H) [M + H] 322.0 A 57 ##STR00377##
4-Chloro-N.sup.3-(3- chloro-4- fluorophenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.37 (s, 1H), 8.03 (s,
1H), 7.25 (bs, 2H), 7.11 (t, J = 9.1 Hz, 1H), 7.06 (s, 1H),
6.59-6.57 (m, 1H), 6.47-6.45 (m, 1H) [M + H] 312.0 A 58
##STR00378## N.sup.3-(3-Bromo-4- fluorophenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.06
(d, J = 5.0 Hz, 1H), 7.11-7.07 (m, 2H), 6.91 (bs, 2H), 6.86 (d, J =
5.0 Hz, 1H), 6.70 (dd, J' = 5.8 Hz, J'' = 2.7 Hz, 1H), 6.50 (dt, J'
= 8.8 Hz, J'' = 6.8 [M + H] 322.0 A Hz, J''' = 3.3 Hz, 1H). 59
##STR00379## N.sup.3-(3,4- Difluoro- phenyl)furo [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.05 (d,
J = 5.0 Hz, 1H), 7.16-7.10 (m, 2H), 6.91 (bs, 2H), 6.85 (d, J = 4.9
Hz, 1H), 6.42-6.37 (m, 1H), 6.30 (d, J = 8.5 Hz, 1H) [-H] 260.0 A
60 ##STR00380## N.sup.3-(4-Fluoro-3- (trifluoro- methyl)phenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.06 (d, J = 5.0 Hz, 1H), 7.29 (s, 1H), 7.21 (t, J = 9.76 Hz,
1H), 6.95 (bs, 2H), 6.88 (d, J = 5.0 Hz, 1H), 6.80-6.78 (m, 1H),
6.73-6.70 (m, 1H) [M + H] 312.0 A 61 ##STR00381## N.sup.3-(3-(tert-
Butyl)phenyl) furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.40 (s, 1H), 8.03 (d, J = 5.0 Hz, 1H), 6.98 (t, J = 7.8
Hz, 1H), 6.83- 6.78 (m, 4H), 6.67- 6.63 (m, 2H), 6.25- 6.23 (m,
1H). 1.20 (s, 9H) [M + H] 282.0 A 62 ##STR00382##
N.sup.3-(4-Fluoro-3- (trifluoro- methoxy) phenyl)furo
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H),
8.03 (d, J = 5.1 Hz, 1H), 7.22-7.17 (m, 2H), 6.96 (bs, 2H), 6.85
(d, J = 5.1 Hz, 1H), 6.55 (d, J = 5.4 Hz, 1H), 6.47 (dt, J' = 8.8
Hz, J'' = 6.5 Hz, J''' = 3.3 Hz, 1H) [M + H] 328.2 A 63
##STR00383## N.sup.3-(5,6,7,8- Tetrahydro- naphthalen-1-
yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.40 (s,
1H), 8.03 (d, J = 5.1 Hz, 1H), 6.81 (d, J = 5.1 Hz, 1H), 6.76- 6.74
(m, 3H), 6.37 (d, J = 7.4 Hz, 1H), 6.01-5.99 (m, 2H), 2.67 (t, J =
5.7 Hz, 2H), 2.61 (t, J = 6.2 Hz, 2H), 1.82-1.81 (m, 2H), 1.72-1.70
[M + H] 280.0 A (m, 2H) 64 ##STR00384## N.sup.3-(3-Chloro-4-
fluorophenyl)- 5- methoxyfuro [2,3-c]pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 7.97 (s, 1H), 7.12 (t, J = 9.1 Hz, 1H), 7.02
(s, 1H), 6.93 (bs, 2H), 6.55 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H),
6.47-6.44 (m, 1H). 6.06 (s, 1H), 3.75 (s, 3H) [M + H] 308.2 A 65
##STR00385## N.sup.3-(3- (Difluoro- methyl) phenyl)furo
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.43 (s, 1H),
8.12 (d, J = 5.1 Hz, 1H), 7.24 (t, J = 7.8 Hz, 1H), 6.95 (d, J =
5.0 Hz, 1H), 7.86 (d, J = 7.4 Hz, 1H), 6.76-6.73 (m, 2H), 6.62 (s,
1H), 5.81 (s, 1H), 5.38 (s, 2H) [M + H] 276.0 A 66 ##STR00386##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-methylfuro [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 7.93 (d, J = 5.1 Hz,
1H), 7.13-7.08 (m, 2H), 6.82 (bs, 2H), 6.71 (d, J = 5.1 Hz, 1H),
6.54 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.48-6.45 (m, 1H), 2.49
(d, J = 2.3 Hz, 3H) [M + H] 292.0 A 67 ##STR00387##
N.sup.3-(3-Chloro-4- fluorophenyl)- 4-methoxyfuro [2,3-c]pyridine-
2,3-diamine DMSO-d.sub.6: .delta. 8.18 (s, 1H), 7.86 (s, 1H), 7.09
(t, J = 9.1 Hz, 1H), 6.99 (s, 1H), 6.68 (bs, 2H), 6.54 (dd, J' =
6.1 Hz, J'' = 2.5 Hz, 1H), 6.46- 6.44 (m, 1H), 3.68 (s, 3H) [M + H]
308.0 A 68 ##STR00388## N.sup.3-(3- Ethynylphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.41 (s, 1H), 8.05 (d,
J = 5.0 Hz, 1H), 7.10-7.06 (m, 2H), 6.89 (bs, 2H), 6.84 (d, J = 5.0
Hz, 1H), 6.70 (d, J = 7.4 Hz, 1H), 6.58- 6.56 (m, 2H), 3.99 (s, 1H)
[M + H] 250.0 A 69 ##STR00389## N.sup.3-(3-(1,1- Difluoroethyl)-
4-fluorophenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.41 (s, 1H), 8.05 (d, J = 5.0 Hz, 1H), 7.09-7.03 (m, 2H),
6.92 (bs, 2H), 6.86 (d, J = 5.0 Hz, 1H), 6.72 (dd, J' = 6.3 Hz, J''
= 2.9 Hz, 1H), 6.55-6.52 (m, 1H), 1.98-1.88 [M + H] 308.2 A (m, 3H)
70 ##STR00390## N.sup.3-(4-Chloro-3- methylphenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.40 (s, 1H), 8.04 (d,
J = 5.0 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.95 (s, 1H), 6.84-6.82
(m, 3H), 6.47 (d, J = 2.4 Hz, 1H), 6.35 (dd, J' = 8.6 Hz, J'' = 2.6
Hz, 1H), 2.17 (s, 3H) [M + H] 274.0 A 71 ##STR00391##
N.sup.3-(5,6,7,8- Tetrahydro- naphthalen-2- yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.38 (s, 1H), 8.02
(d, J = 5.0 Hz, 1H), 6.81 (d, J = 5.0 Hz, 1H), 6.76- 6.72 (m, 3H),
6.55 (s, 1H), 6.29 (dd, J' = 8.1 Hz, J'' = 2.0 Hz, 1H), 6.20 (bs,
1H), 2.55-2.50 (m, 4H), [M + H] 280.0 A 1.65 (s, 4H) 72
##STR00392## N.sup.3-(3-Chloro-4- fluorophenyl)- 4,7-dimethyl-
furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.67 (s,
1H), 7.08 (t, J = 9.0 Hz, 1H), 7.03 (s, 1H), 6.73 (bs, 2H),
6.50-6.48 (m, 1H), 6.42-6.40 (m, 1H), 2.41 (s, 3H), 2.06 (s, 3H)
HRMS [M + H] 306.08 0 A 73 ##STR00393## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-ethylfuro[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 7.97 (d, J = 5.1 Hz, 1H), 7.14-7.08 (m, 2H),
6.81 (bs, 2H), 6.72 (d, J = 5.1 Hz, 1H), 6.55 (dd, J' = 6.3 Hz, J''
= 2.7 Hz, 1H), 6.48-6.44 (m, 1H), 2.86 (q, J = 7.6 Hz, 2H), 1.28
(t, J = 7.6 Hz, 3H) [M + H] 306.1 A 74 ##STR00394##
7-Ethyl-N.sup.3-(4- fluoro-3- (trifluoro- methyl)phenyl)
furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.97 (d, J
= 5.0 Hz, 1H), 7.27 (s, 1H), 7.20 (t, J = 9.8 Hz, 1H), 6.84 (s,
2H), 6.80-6.78 (m, 1H), 6.73-6.69 (m, 2H), 2.86 (q, J = 7.5 Hz,
2H), 1.28 (t, J = 7.5 Hz, 3H) [M - H] 338.0 A 75 ##STR00395##
N.sup.3-(3- Benzylphenyl) furo[2,3-c] pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.41 (s, 1H), 8.09 (s, 1H), 7.24-7.18 (m, 5H),
7.03 (s, 1H), 6.89 (s, 1H), 6.57-6.43 (m, 3H), 5.54 (s, 1H), 5.29
(s, 2H), 3.81 (s, 2H) [M + H] 316.2 A 76 ##STR00396##
7-Ethyl-N.sup.3-(3- (trifluoro- methyl)phenyl) furo[2,3-c]
pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.97 (d, J = 5.1 Hz,
1H), 7.35 (s, 1H), 7.28 (t, J = 7.9 Hz, 1H), 6.89 (d, J = 7.6 Hz,
1H), 6.83-6.80 (m, 3H), 6.74-6.71 (m, 2H), 2.86 (q, J = 7.6 Hz,
2H), 1.29 (t, J = 7.6 Hz, 3H) [M - H] 320.0 A 77 ##STR00397##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-propylfuro [2,3-c]pyridine-
2,3-diamine DMSO-d.sub.6: .delta. 7.96 (d, J = 5.1 Hz, 1H),
7.14-7.07 (m, 2H), 6.81 (bs, 2H), 6.71 (d, J = 5.1 Hz, 1H), 6.55
(dd, J' = 6.3 Hz, J'' = 2.7 Hz, 1H), 6.47-6.44 (m, 1H), 2.81 (t, J
= 7.6 Hz, 2H), 1.79-1.71 (m, 2H), 0.94 (t, J = 7.4 Hz, 3H) [M + H]
320.0 A 78 ##STR00398## 7-Propyl-N.sup.3-(3- (trifluoro-
methyl)phenyl) furo[2,3-c] pyridine- 2,3-diamine CD.sub.3CN:
.delta. 8.04 (d, J = 5.1 Hz, 1H), 7.29 (t, J = 8.2 Hz, 1H), 6.97
(d, J = 7.6 Hz, 1H), 6.83- 6.80 (m, 3H), 5.91 (s, 1H), 5.30 (s,
2H), 2.89 (t, J = 7.6 Hz, 2H), 1.87-1.78 (m, 2H), 0.98 (t, J = 7.4
Hz, 3H) [M + H] 336.0 A 79 ##STR00399## N.sup.3-(4-Fluoro-3-
(trifluoro- methyl)phenyl)- 7-propylfuro [2,3-c]pyridine-
2,3-diamine DMSO-d.sub.6: .delta. 7.97 (d, J = 4.8 Hz, 1H), 7.27
(s, 1H), 7.20 (t, J = 9.7 Hz, 1H), 6.85 (s, 2H), 6.80 (s, 1H),
6.73-6.71 (m, 2H), 2.81 (t, J = 7.3 Hz, 2H), 1.79-1.73 (m, 2H),
0.94 (t, J = 7.2 Hz, 3H) [M + H] 354.4 A 80 ##STR00400##
7-Methyl-N.sup.3- (3-(trifluoro- methyl)phenyl) furo[2,3-c]
pyridine- 2,3-diamine CD.sub.3CN: .delta. 8.00 (s, 1H), 7.30-7.27
(m, 1H), 6.97 (d, J = 6.8 Hz, 1H), 6.82 (bs, 3H), 5.90 (s, 1H),
5.32 (s, 2H), 2.55 (s, 3H) [M + H] 308.2 A 81 ##STR00401##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-phenylfuro [2,3-c]pyridine-
2,3-diamine CD.sub.3CN: .delta. 8.32-8.30 (m, 2H), 8.22 (d, J = 5.0
Hz, 1H), 7.56- 7.52 (m, 2H), 7.48- 7.44 (m, 1H), 7.03 (t, J = 9.1
Hz, 1H), 6.95 (d, J = 5.0 Hz, 1H), 6.65 (dd, J' = 6.3 Hz, J'' = 2.8
Hz, 1H), 6.60-6.56 (m, 1H), 5.71 (s, 1H), 5.42 (s, 2H) [M + H]
354.0 A 82 ##STR00402## 7-Phenyl-N.sup.3- (3-(trifluoro-
methyl)phenyl) furo[2,3-c] pyridine-2,3- diamine CD.sub.3CN:
.delta. 8.31 (d, J = 7.5 Hz, 2H), 8.22 (d, J = 4.9 Hz, 1H), 7.55
(t, J = 7.5 Hz, 2H), 7.46 (t, J = 7.2 Hz, 1H), 7.31 (t, J = 8.0 Hz,
1H), 6.99 (d, J = 7.5 Hz, 1H), 6.95 (d, J = 5.0 Hz, 1H), 6.88 (bs,
2H), 5.96 (s, 1H), 5.47 (s, 2H) [M + H] 370.0 A 83 ##STR00403##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-isopropyl- furo[2,3-c]
pyridine- 2,3-diamine CD.sub.3CN: .delta. 8.05 (d, J = 5.0 Hz, 1H),
7.01 (t, J = 9.1 Hz, 1H), 6.80 (d, J = 5.0 Hz, 1H), 6.61 (dd, J' =
6.3 Hz, J'' = 2.8 Hz, 1H), 6.56-6.52 (m, 1H), 5.66 (s, 1H), 5.26
(s, 2H), 3.46-3.39 (m, 1H), 1.34 (d, J = 6.9 Hz, 6H) [M + H] 320.0
A 84 ##STR00404## N.sup.3-(3- (Cyclopropyl- methyl)phenyl)
furo[2,3-c] pyridine-2,3- diamine CD.sub.3CN: .delta. 8.42 (s, 1H),
8.11 (d, J = 5.1 Hz, 1H), 7.03 (t, J = 7.8 Hz, 1H), 6.94 (d, J =
5.0 Hz, 1H), 6.61 (d, J = 7.5 Hz, 1H), 6.53 (s, 1H), 6.43 (dd, J' =
8.0 Hz, J'' = 1.6 Hz, 1H), [M + H] 280.0 A 5.52 (s, 1H), 5.30 (s,
2H), 2.38 (d, J = 6.9 Hz, 2H), 0.93- 0.86 (m, 1H), 0.44- 0.40 (m,
2H), 0.14- 0.12 (m, 2H) 85 ##STR00405## N.sup.3-(3-Chloro-4-
fluorophenyl)- 5-methoxy-7- methylfuro[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 7.11 (t, J = 9.1 Hz, 1H), 7.01 (s, 1H), 6.84
(bs, 2H), 6.53 (dd, J' = 6.3 Hz, J'' = 2.8 Hz, 1H), 6.47-6.43 (m,
1H), 5.92 (s, 1H), 3.73 (s, 3H), 2.40 (s, 3H) [M - H] 320.0 A 86
##STR00406## 7-Benzyl-N.sup.3-(3- chloro-4- fluorophenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 7.96 (d, J
= 5.1 Hz, 1H), 7.32-7.30 (m, 4H), 7.24-7.22 (m, 1H), 7.11-7.06 (m,
2H), 6.77 (bs, 2H), 6.65 (d, J = 5.1 Hz, 1H), 6.56 (dd, J' = 6.2
Hz, J'' = 2.5 Hz, 1H), 6.50-6.48 (m, 1H), 4.08 (s, 2H) [M + H]
368.0 A 87 ##STR00407## N.sup.3-(3-Chloro-5- methylphenyl)
furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.42 (s,
1H), 8.06 (d, J = 5.0 Hz, 1H), 7.12 (s, 1H), 6.98 (bs, 2H), 6.87
(d, J = 5.0 Hz, 1H), 6.45 (s, 1H), 6.29 (s, 2H), 2.14 (s, 3H) [M +
H] 274.0 A
88 ##STR00408## N.sup.3-(3- Cyclohexyl- phenyl)furo
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.39 (s, 1H),
8.03 (d, J = 5.1 Hz, 1H), 6.96 (t, J = 7.8 Hz, 1H), 6.83 (d, J =
5.1 Hz, 1H), 6.77 (d, J = 9.1 Hz, 3H), 6.46 (d, J = 7.6 Hz, 1H),
6.42 (s, 1H), 6.30 (dd, J'= 8.0 Hz, [M + H] 308.0 A J'' = 1.5 Hz,
1H), 2.31 (bs, 1H), 173- 1.64 (m, 5H), 1.35- 1.24 (m, 4H), 1.20-
1.15 (m, 1H) 89 ##STR00409## N.sup.3-(3- (Cyclobutyl-
methyl)phenyl) furo[2,3-c] pyridine- 2,3-diamine CDCl.sub.3:
.delta. 8.52 (s, 1H), 8.20 (d, 1H, J = 5 Hz), 7.08-7.02 (m, 2H),
6.58 (d, 1H, J = 7.2 Hz), 6.45 (d, 1H, J = 7.8 Hz), 6.41 (s, 1H),
4.71 (bs, 1H), 4.51 (bs, 2H), 2.59- 2.57 (m, 2H), 2.50- [M + H]
294.4 A 2.45 (m, 1H), 1.98 (m, 2H), 1.82-1.76 (m, 2H), 1.70-1.65
(m, 2H) 90 ##STR00410## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(3-
methoxyphenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.16 (d, 1H, J = 5.0 Hz), 7.87 (d, 1H, J = 7.8 Hz), 7.84
(m, 1H), 7.44 (t, 1H, J = 8.0 Hz), 7.15-7.11 (m, 2H), 7.03-7.01 (m,
3H), 6.85 (d, 1H, J = 5.0 Hz), 6.60-6.58 (m, 1H), 6.49 (m, 1H),
3.85 (s, 3H) [M + H] 384.4 A 91 ##STR00411## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(4- chlorophenyl) furo[2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.30 (d, 2H, J = 8.3 Hz), 8.16 (d,
1H, J = 4.9 Hz), 7.60 (d, 2H, J = 8.3 Hz), 7.14-7.11 (m, 2H), 7.04
(s, 2H), 6.87 (d, 1H, J = 4.9 Hz), 6.60 (m, 1H), 6.52 (m, 1H) [M +
H] 388.1 A 92 ##STR00412## N.sup.3-(3- Ethynylphenyl)- 7-
phenylfuro [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.27
(d, 2H, J = 7.5 Hz), 8.16 (d, 1H, J = 4.8 Hz), 7.54 (t, 2H, J = 7.5
Hz), 7.47-7.45 (m, 1H), 7.11-7.08 (m, 2H), 6.99 (bs, 2H), 6.84 (d,
1H, J = 4.8 Hz), 6.71 (d, 1H, J = 7.2 Hz), 6.63- 6.60 (m, 2H), 3.99
(s, 1H) [M + H] 326.0 A 93 ##STR00413## N.sup.3-(3- ethynylphenyl)-
7- methylfuro [2,3-c] pyridine-2,3- diamine CD.sub.3CN: .delta. 8.0
(d, 1H, J = 5.04 Hz), 7.11 (t, 1H, J = 8.3 Hz), 6.82-6.79 (m, 2H),
6.64 (m, 2H), 5.68 (s, 1H), 5.27 (s, 2H), 3.52 (s, 1H), 2.54 (s,
3H) [M + H] 264.0 A 94 ##STR00414## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(pyridin-3- yl)furo[2,3- c]pyridine-2,3- diamine
CD.sub.3CN: .delta. 9.51 (s, 1H), 8.63 (m, 2H), 8.25 (m, 1H), 7.51
(m, 1H), 7.03-6.99 (m, 2H), 6.66-6.59 (m, 2H), 5.73 (s, 1H), 5.51
(s, 2H) [M + H] 355.0 A 95 ##STR00415## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(2- chlorophenyl) furo[2,3- e]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.13 (d, J = 5.1 Hz, 1H), 7.61-7.55
(m, 2H), 7.50-7.47 (m, 2H), 7.17-7.13 (m, 2H), 6.95 (bs, 2H), 6.91
(d, J = 5.1 Hz, 1H), 6.59 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H),
6.51-6.47 (m, 1H) [M + H] 388 A 96 ##STR00416## 7-Ethyl-N.sup.3-(3-
ethynylphenyl) furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 7.96 (s, 1H), 7.04 (d, J = 16.0 Hz, 2H), 6.77-6.69 (m, 3H),
6.55 (bs, 2H), 3.98 (s, 1H), 2.86 (s, 2H), 1.28 (s, 3H) [M + H]
278.1 A 97 ##STR00417## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.73 (d, J = 6.1 Hz, 2H), 8.27-8.26 (m, 3H), 7.05-7.01 (m,
2H), 6.66 (dd, J' = 6.2 Hz, J'' = 2.8 Hz, 1H), 6.60-6.56 (m, 1H),
5.73 (s, 1H), 5.54 (bs, 2H) [M + H] 355.2 A 98 ##STR00418##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(2- methoxyphenyl) furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.09 (d, J = 5.0 Hz,
1H), 7.44-7.37 (m, 2H), 7.17-7.14 (m, 2H), 7.12-7.04 (m, 2H),
6.85-6.82 (m, 3H), 6.58 (dd, J' = 6.2 Hz, J'' = 2.5 Hz, 1H),
6.50-6.48 (m, 1H) [M + H] 384.2 A 99 ##STR00419## 3-((2- Aminofuro
[2,3-c] pyridin-3- yl)amino) benzonitrile CD.sub.3CN: .delta. 8.44
(s, 1H), 8.12 (d, J = 4.6 Hz, 1H), 7.27 (t, J = 7.6 Hz, 1H), 7.02
(d, J = 6.9 Hz, 1H), 6.95-6.86 (m, 3H), 5.94 (s, 1H), 5.39 (s, 2H).
[M + H] 251.2 A 100 ##STR00420## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(3- chlorophenyl) furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.28- 8.25 (m, 2H), 8.18 (d, J = 4.9
Hz, 1H), 7.59-7.51 (m, 2H), 7.16-7.09 (m, 4H), 6.88 (d, J = 4.9 Hz,
1H), 6.60-6.59 (m, 1H), 6.52-6.50 (m, 1H) [M + H] 388 A 101
##STR00421## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(4-
methoxyphenyl) furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.24 (d, J = 8.7 Hz, 2H), 8.13 (d, J = 5.0 Hz, 1H),
7.15-7.07 (m, 4H), 6.95 (bs, 2H), 6.80 (d, J = 5.0 Hz, 1H), 6.58
(dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.52-6.49 (m, 1H), 3.84 (s,
3H) [M + H] 384 A 102 ##STR00422## 5-Butoxy-N.sup.3- (3-chloro-4-
fluorophenyl)- 7-phenylfuro [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.27 (d, J = 7.5 Hz, 2H), 7.52 (t, J = 7.5
Hz, 2H), 7.43 (t, J = 7.5 Hz, 1H), 7.13 (t, J = 9.1 Hz, 1H), 7.06
(s, 1H), 7.02 (bs, 2H), 6.59 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H),
6.52-6.49 (m, 1H), 6.04 (s, 1H), 4.28 (t, J = 6.5 Hz, 2H),
1.72-1.65 (m, 2H), 1.46-1.37 (m, 2H), 0.92 (t, J = 7.4 Hz, 3H) [M +
H] 426.2 C 103 ##STR00423## 2-Amino-3-((3- chloro-4- fluorophenyl)
amino)furo [2,3-c] pyridine-7- carbonitrile CD.sub.3CN: .delta.
8.18 (d, J = 5.1 Hz, 1H), 7.15 (d, J = 5.0 Hz, 1H), 7.03 (t, J =
9.0 Hz, 1H), 6.64-6.62 (m, 1H), 6.56-6.52 (m, 1H), 5.81 (bs, 2H),
5.71 (s, 1H) [M + H] 303 A 104 ##STR00424## N.sup.3-(3-Chloro-4-
fluorophenyl)- 5-phenoxy-7- phenylfuro[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.11 (d, J = 6.6 Hz, 2H), 7.47-7.37 (m, 5H),
7.15-7.11 (m, 7H), 6.60 (bs, 1H), 6.52 (bs, 1H), 6.26 (s, 1H) [M +
H] 444.2 C 105 ##STR00425## N.sup.3-(3-Chloro-4- fluorophenyl)-
5-ethoxy-7- phenylfuro[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.27 (d, J = 7.5 Hz, 2H), 7.52 (t, J = 7.4 Hz, 2H),
7.45-7.42 (m, 1H), 7.13 (t, J = 9.1 Hz, 1H), 7.06 (s, 1H), 7.01 (s,
2H), 6.60 (bs, 1H), 6.51-6.49 (m, 1H), 6.03 (s, 1H). 4.34 (q, J =
6.8 Hz, 2H), 1.31 (t, J = 6.9 Hz, 3H) [M + H] 398.2 C 106
##STR00426## 5-(tert-Butoxy)- N.sup.3-(3-chloro-4- fluorophenyl)-
7-phenylfuro [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.24 (d, J = 7.8 Hz, 2H), 7.54 (t, J = 7.4 Hz, 2H), 7.43 (d, J =
7.4 Hz, 1H), 7.13 (t, J = 9.2 Hz, 1H), 7.04 (s, 1H), 6.97 (bs, 2H),
6.60-6.59 (m, 1H), 6.51-6.49 (m, 1H), 5.99 (s, 1H), 1.54 (s, 9H) [M
+ H] 426.2 A 107 ##STR00427## N.sup.3-(3-Chloro-4- fluorophenyl)-
5-isopropoxy-7- phenylfuro[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.26 (d, J = 7.7 Hz, 2H), 7.52 (t, J = 7.5
Hz, 2H), 7.43 (t, J = 7.1 Hz, 1H), 7.13 (t, J = 9.1 Hz, 1H), 7.05
(s, 1H), 7.02 (bs, 2H), 6.60 (d, J = 3.9 Hz, 1H), 6.51-6.48 (m,
1H), 5.98 (s, 1H), 5.32-5.26 (m, 1H), 1.29 (d, J = 6.0 Hz, 6H) [M +
H] 412 A 108 ##STR00428## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-cyclohexyl- furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6:
.delta. 8.03 (d, J = 5.0 Hz, 1H), 7.01 (t, J = 9.1 Hz, 1H), 6.78
(d, J = 5.0 Hz, 1H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.7 Hz, 1H),
6.55-6.52 (m, 1H), 5.67 (s, 1H), 5.27 (bs, 2H), 3.09 (t, J = 11.6
Hz, 1H), 1.87-1.81 (m, 4H), 1.78-1.73 (m, 3H), 1.51-1.42 (m, 2H),
1.38-1.27 (m, 1H) [M + H] 360 A 109 ##STR00429##
N.sup.3-(3-Chloro-4- fluorophenyl)- 5-ethyl-7- phenylfuro[2,3-
c]pyridine-2,3- diamine CDCl.sub.3: .delta. 8.24 (d, J = 7.2 Hz,
2H), 7.50 (t, J = 7.1 Hz, 2H), 7.43-7.40 (m, 1H), 6.96 (t, J = 8.6
Hz, 1H), 6.81 (s, 1H), 6.65 (bs 1H), 6.51 (bs, 1H), 4.74 (s, 1H),
4.49 (s, 2H), 2.84 (q, J = 7.7 Hz, 2H), 1.32 (t, J = 7.5 Hz, 3H) [M
+ H] 382 A 110 ##STR00430## N.sup.3-(3-Ethynyl- 4- fluorophenyl)-
7- phenylfuro[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6: .delta.
8.27 (d, J = 7.4 Hz, 2H), 8.16 (d, J = 5.1 Hz, 1H), 7.53 (t, J =
7.5 Hz, 2H), 7.45 (t, J = 7.3 Hz, 1H), 7.06-7.03 (m, 2H), 7.01-7.00
(m, 2H), 6.85 (d, J = 5.0 Hz, 1H), 6.62- 6.56 (m, 2H), 4.33 (s, 1H)
[M + H] 344 A 111 ##STR00431## N.sup.3-(3-Ethynyl- 4-
fluorophenyl)- 7-(pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.74 (d, J = 5.6 Hz, 2H), 8.23-8.21 (m, 3H),
7.12 (bs, 2H), 7.05- 7.01 (m, 2H), 6.94 (d, J = 5.1 Hz, 1H),
6.62-6.57 (m, 2H), 4.31 (s, 1H) [M + H] 345.2 A 112 ##STR00432##
2-Amino-3-((3- chloro-4- fluorophenyl) amino)benzo- furan-6-
carbonitrile DMSO-d.sub.6: .delta. 7.75 (s, 1H), 7.39 (d, J = 8.1
Hz, 1H), 7.14-7.10 (m, 2H), 6.96 (bs, 2H), 6.92 (d, J = 8.0 Hz,
1H), 6.56 (dd, J' = 6.3 Hz, J'' = 2.7 Hz, 1H), 6.47 (dt, J' = 8.8
Hz, J'' = 6.6 [M - H] 300 A, B Hz, J''' = 3.2 Hz, 1H) 113
##STR00433## 2-Amino-3-((3- (trifluoro- methyl)phenyl) amino)
benzofuran-6- carbonitrile CD.sub.3CN: .delta. 7.60 (s, 1H), 7.39
(d, J = 7.4 Hz, 1H), 7.29 (s, 1H), 7.05 (d, J = 7.6 Hz, 1H), 6.98
(d, J = 6.8 Hz, 1H), 6.84 (bs, 2H), 5.93 (s, 1H), 5.39 (bs, 2H) [M
- H] 316 A, B 114 ##STR00434## 2-Amino-3-(m- tolylamino)
benzofuran-6- carbonitrile DMSO-d.sub.6: .delta. 7.73 (s, 1H), 7.37
(d, J = 7.9 Hz, 1H), 6.95 (t, J = 7.8 Hz, 1H), 6.89 (d, J = 8.0 Hz,
1H), 6.83 (s, 2H), 6.80 (s, 1H), 6.42 (d, J = 7.4 Hz, 1H),
6.35-6.31 (m, 2H), 2.14 (s, 3H) [M - H] 262 A, B 115 ##STR00435##
2-Amino-3-((3- bromophenyl) amino) benzofuran-6- carbonitrile
CD.sub.3CN: .delta. 7.59 (s, 1H), 7.38 (dd, J' = 8.1 Hz, J'' = 1.1
Hz, 1H), 7.06-7.02 (m, 2H), 6.83 (dd, J' = 7.8 Hz, J'' = 0.9 Hz,
1H), 6.72 (t, J = 2.0 Hz, 1H), 6.60 (dd, J' = 8.2 Hz, J'' = 2.2 [M
- H] 326 A, B Hz, 1H), 5.78 (s, 1H), 5.37 (bs, 2H) 116 ##STR00436##
2-Amino-3-((2- chlorophenyl) amino) benzofuran-6- carbonitrile
CD.sub.3CN: .delta. 7.60 (s, 1H), 7.36 (dd, J' = 22.0 Hz, J'' = 7.8
Hz, 2H), 7.03-7.02 (m, 2H), 6.69 (t, J = 7.2 Hz, 1H), 6.42 (d, J =
7.9 Hz, 1H), 5.82 (s, 1H), 5.42 (bs, 2H) [M - H] 282 A, B 117
##STR00437## N.sup.3-(3-Chloro-4- fluorophenyl) benzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.56 (d, J
= 7.5 Hz, 1H), 7.22 (s, 1H), 7.11-7.06 (m, 2H), 6.97-6.94 (m, 2H),
6.48-6.46 (m, 1H), 6.44-6.41 (m, 1H), 5.85 (bs, 2H) [M + H] 293.0
B, E 118 ##STR00438## N.sup.3- Phenylbenzo[b] thiophene-2,3-
diamine (500 MHz, DMSO- d.sub.6): .delta. 7.55 (d, J = 7.5 Hz, 1H),
7.07 (t, J = 7.2 Hz, 1H), 7.03- 6.98 (m, 3H), 6.94 (t, J = 7.2 Hz,
2H), 6.53 (t, J = 7.3 Hz, 1H), 6.47 (d, J = 7.6 Hz, 2H), 5.70 (bs,
2H) [M + H] 240.1 B, E 119 ##STR00439## N.sup.3-(3- (Trifluoro-
methyl)phenyl) benzo[b] thiophene- 2,3-diamine (500 MHz, DMSO-
d.sub.6): .delta. 7.57 (d, J = 8.0 Hz, 1H), 7.49 (s, 1H), 7.23 (t,
J = 8.0 Hz, 1H), 7.09 (t, J = 7.0 Hz, 1H), 6.97- 6.93 (m, 2H), 6.83
(d, J = 7.3 Hz, 1H), 6.76 (bs, 1H), 6.67 [M + H] 309.1 B, E (d, J =
8.0 Hz, 1H), 5.87 (bs, 2H) 120 ##STR00440## N.sup.3-(3-
Fluorophenyl) benzo[b] thiophene-2,3- diamine CDCl.sub.3: .delta.
7.59 (d, J = 7.8 Hz, 1H), 7.25-7.21 (m, 2H), 7.15-7.06 (m, 2H),
6.46-6.41 (m, 2H), 6.32-6.28 (m, 1H), 5.08 (s, 1H), 4.14 (bs, 2H)
[M + H] 259.1 B, E 121 ##STR00441## N.sup.3-(3-Chloro-4-
fluorophenyl) thieno[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.66 (s, 1H), 8.13 (d, J = 5.4 Hz, 1H), 7.27 (s, 1H), 7.11
(t, J = 9.1 Hz, 1H), 6.86 (d, J = 5.3 Hz, 1H), 6.72 (bs, 2H), 6.50
(dd, J' = 6.2 Hz, J'' = 2.4 Hz, 1H), 6.42-6.40 (m, [M + H] 294 D
1H) 122 ##STR00442## 3-((3- Chlorophenyl) thio)benzo[b] thiophen-2-
amine (500 MHz, CDCl.sub.3): .delta. 7.61 (d, J = 7.7 Hz, 1H), 7.47
(d, J = 8.0 Hz, 1H), 7.27 (dd, J' = 14.7 Hz, J'' = 7.0 Hz, 1H),
7.16 (t, J = 7.7 Hz, 1H), 7.11- 7.04 (m, 3H), 6.92 (d, J = 7.1 Hz,
1H), [M + H] 292 E, K 4.83 (bs, 2H) 123 ##STR00443##
5-Bromo-N.sup.3-(3- chloro-4- fluorophenyl) benzo[b] thiophene-2,3-
diamine (500 MHz, DMSO- d.sub.6): .delta. 7.53 (d, J = 8.3 Hz, 1H),
7.24 (s, 1H), 7.10 (d, J = 9.2 Hz, 1H), 7.07 (dd, J' = 8.3 Hz, J''
= 1.8 Hz, 1H), 7.03 (d, J = 1.8 Hz, 1H), 6.47 (dd, J' = 6.5 Hz, J''
= 2.8 Hz, 1H), 6.40 (dt, J' = 8.9 Hz, J'' = 3.4 Hz, 1H), 6.14 (bs,
2H) [M + H] 370.9 E 124 ##STR00444## N.sup.3-(4-Chloro-3-
(trifluoro- methyl)phenyl) benzo[b] thiophene-2,3- diamine (500
MHz, DMSO- d.sub.6): .delta. 7.67 (s, 1H), 7.56 (d, J = 7.7 Hz,
1H), 7.30 (d, J = 8.6 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.97- 6.93
(m, 3H), 6.58 (d, J = 7.8 Hz, 1H), 5.94 (bs, 2H) [M + H] 343 E 125
##STR00445## 3-((3- Chlorophenyl) thio)benzo[b] thiophen-2- amine
hydrochloride DMSO-d.sub.6: .delta. 7.65 (d, J = 7.8 Hz, 1H), 7.24
(t, J = 7.9 Hz, 1H), 7.20-7.17 (m, 2H), 7.15-7.12 (m, 1H),
7.05-7.01 (m, 1H), 6.97 (d, J = 7.8 Hz, 1H), 6.94-6.93 (m, 1H),
6.45 (bh, 2H) [M - HCl] 290.9 E, K 126 ##STR00446##
N.sup.3-(4-Bromo-3- (trifluoro- methyl)phenyl) benzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.68 (s,
1H), 7.56 (d, J = 7.8 Hz, 1H), 7.44 (d, J = 8.9 Hz, 1H), 7.10 (dd,
J' = 8.0 Hz, J'' = 0.85 Hz, 1H), 6.97-6.93 (m, 3H), 6.50 (d, J =
7.2 Hz, 1H), 5.94 [M - H] 384.9 E (bs, 2H) 127 ##STR00447## 3-(3-
Chlorophenoxy) benzo[b] thiophen-2- amine hydrochloride
DMSO-d.sub.6: .delta. 7.63 (d, J = 8.1 Hz, 1H), 7.31 (t, J = 8.1
Hz, 1H), 7.13 (t, J = 7.5 Hz, 1H), 7.07-7.00 (m, 2H), 6.92-6.85 (m,
3H), 5.66 (bh, 2H) [M - HCl] 276 E, K 128 ##STR00448##
N.sup.3-(4-Fluoro- 3-(trifluoro- methyl)phenyl) benzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.57
(d, J = 7.5 Hz, 1H), 7.42 (s, 1H), 7.18-7.09 (m, 2H), 6.97-6.94 (m,
2H), 6.75 (dd, J' = 6.0 Hz, J'' = 2.6 Hz, 1H), 6.65-6.62 (m, 1H),
5.89 (bs, 2H) [M + H] 327.1 E 129 ##STR00449## 5-Chloro-N.sup.3-
(3-chloro-4- fluorophenyl) benzo[b] thiophene-2,3- diamine
hydrochloride (500 MHz, DMSO- d.sub.6): .delta. 7.59 (d, J = 8.3
Hz, 1H), 7.09 (t, J = 9.2 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 6.88
(s, 1H), 6.49-6.48 (m, 1H), 6.41-6.39 (m, 1H), 4.67 (bh, 4H) [M +
Cl] 360.9 E 130 ##STR00450## 5-Chloro-N.sup.3- phenylbenzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.58 (d, J
= 8.3 Hz, 1H), 7.03 (t, J = 7.3 Hz, 2H), 6.99 (s, 1H), 6.93 (d, J =
8.3 Hz, 1H), 6.85 (s, 1H), 6.56 (t, J = 7.6 Hz, 1H), 6.46 (d, J =
8.3 Hz, 2H), 6.00 (bs, [M + H] 275 E 2H) 131 ##STR00451##
5-Chloro-N.sup.3-(4- fluorophenyl) benzo[b] thiophene-2,3- diamine
(500 MIlz, DMSO- d.sub.6): .delta. 7.57 (d, J = 8.3 Hz, 1H), 6.97
(s, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.90-6.86 (m, 3H), 6.44 (dd, J'
= 8.6 Hz, J'' = 4.6 Hz, 2H), 6.03 (bs, 2H) [M + H] 293 E 132
##STR00452## 5-Chloro-N.sup.3-(4- fluoro-3- (trifluoro-
methyl)phenyl) benzo[b] thiophene-2,3- diamine (500 MHz, DMSO-
d.sub.6): .delta. 7.59 (d, J = 8.3 Hz, 1H), 7.44 (s, 1H), 7.17 (t,
J = 9.8 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.90 (s, 1H), 6.76-6.75
(m, 1H), 6.63-6.61 (m, 1H), 6.18 (bs, 2H) [M - H] 359 E 133
##STR00453## 5-Chloro-N.sup.3-(3- (trifluoro- methyl)phenyl)
benzo[b] thiophene-2,3- diamine hydrochloride (500 MHz, DMSO-
d.sub.6, salt free): .delta. 7.60 (d, J = 8.2 Hz, 1H), 7.51 (s,
1H), 7.25 (t, J = 7.8 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.88-6.85
(m, 2H), 6.75 (s, 1H), 6.65 (d, J = 8.3 Hz, 1H), 6.17 [M - H] 341 E
(bs, 2H) 134 ##STR00454## N.sup.3-(3-Chloro-4- fluorophenyl)-
5-phenylbenzo [b]thiophene- 2,3-diamine (500 MHz, DMSO- d.sub.6):
.delta. 7.65 (d, J = 8.3 Hz, 1H), 7.50 (d, J = 7.4 Hz, 2H), 7.39-
7.35 (m, 3H), 7.28- 7.23 (m, 2H), 7.18 (s, 1H), 7.07 (t, J = 9.0
Hz, 1H), 6.54-6.52 (m, 1H), 6.47-6.44 [M - H] 367 E (m, 1H), 5.94
(bs, 2H) 135 ##STR00455## N.sup.3-Methyl-N.sup.3- phenylbenzo
[b]thiophene- 2,3-diamine hydrochloride DMSO-d.sub.6, salt free:
.delta. 7.58 (d, J = 9.8 Hz, 1H), 7.12-7.03 (m, 3H), 6.95 (t, J =
7.6 Hz, 1H), 6,76 (dd, J' = 7.8 Hz, J'' = 0.5 Hz, 1H), 6.60 (td, J'
= 8.3 Hz, J'' = 1.0 Hz, 1H), 6.54-6.51 [M + H] 255.1 E (m, 2H),
5.99 (bs, 2H), 3.18 (s, 3H) 136 ##STR00456## 2-Amino-3-((3-
chloro-4- fluorophenyl) amino)benzo [b]thiophene- 6-carbonitrile
DMSO-d.sub.6: .delta. 8.11 (s, 1H), 7.47 (d, J = 8.3 Hz, 1H), 7.32
(s, 1H), 7.11 (t, J = 9.1 Hz, 1H), 7.01 (d, J = 8.2 Hz, 1H), 6.63
(bs, 2H), 6.51 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.43-6.40 (m,
1H) [M - H] 316 E, F 137 ##STR00457## N.sup.3-(3- Ethynyl-
phenyl)-1- benzothio- phene-2,3- diamine (500 MHz, DMSO- d.sub.6):
.delta. 7.56 (d, J = 7.7 Hz, 1H), 7.19 (s, 1H), 7.08 (d, J = 7.7
Hz, 1H), 7.05-7.02 (m, 1H), 6.94 (dd, J' = 13.1 Hz, J'' = 7.3 Hz,
2H), 6.64 (d, J = 7.4 Hz, 1H), 6.54 (d, [M + H] 265.1 E, F J = 8.0
Hz, 1H), 6.47 (s, 1H), 5.80 (bs, 2H), 3.93 (s, 1H) 138 ##STR00458##
N.sup.3-(2,3- Dimethyl- phenyl)-1- benzothio- phene-2,3- diamine
(500 MHz, DMSO- d.sub.6): .delta. 7.55 (d, J = 7.6 Hz, 1H), 7.05
(t, J = 7.3 Hz, 1H), 6.93 (t, J = 6.9 Hz, 1H), 6.86 (d, J = 7.6 Hz,
1H), 6.66 (t, J = 7.6 Hz, 1H), 6.42 (d, J = 7.4 Hz, 1H), 6.19 (s,
1H), 5.91 (d, J = 7.9 Hz, 1H), 5.69 [M + H] 269.1 E (bs, 2H), 2.19
(d, J = 4.3 Hz, 6H) 139 ##STR00459## Benzyl (3-((3- chloro-4-
fluorophenyl) amino)furo [2,3-c]pyridin- 2-yl)carbamate
DMSO-d.sub.6: .delta. 10.39 (s, 1H), 8.85 (s, 1H), 8.35 (d, J = 8.5
Hz, 1H), 7.80 (s, 1H), 7.33 (bs, 6H), 7.15 (t, J = 9.0 Hz, 1H),
6.73 (d, J = 3.2 Hz, 1H), 6.61 (s, 1H), 5.12 (s, 2H) [M - H] 410 G
140 ##STR00460## Benzyl (2- (((benzyloxy) carbonyl) amino)furo
[2,3-c]pyridin- 3-yl)(3-chloro- 4-fluorophenyl) carbamate
DMSO-d.sub.6: .delta. 8.96 (s, 1H), 8.39-8.36 (m, 2H), 7.45-7.15
(m, 12 H), 6.75 (s, 1H), 6.62 (s, 1H), 5.20 (s, 4H) [M + H] 546 H
141 ##STR00461## Ethyl (3-((3- chloro-4- fluorophenyl) amino)furo
[2,3-c]pyridin- 2-yl)carbamate DMSO-d.sub.6: .delta. 10.19 (s, 1H),
8.83 (s, 1H), 8.34 (d, J = 5.0 Hz, 1H), 7.75 (s, 1H), 7.31 (d, J =
5.2 Hz, 1H), 7.18 (t, J = 9.1 hz, 1H), 6.73-6.71 (m, 1H), 6.63-6.60
(m, 1H), 4.07 (q, J = 7.0 Hz, 2H), 1.14 (t, J = 7.0 Hz, 3H) [M + H]
350 G 142 ##STR00462## Ethyl (3- chloro-4- fluorophenyl)
(2-((ethoxy- carbonyl) amino)furo [2,3-c]pyridin- 3-yl)carbamate
DMSO-d.sub.6: .delta. 8.97 (s, 1H), 8.42 (d, J = 5.2 Hz, 1H), 8.28
(s, 1H), 7.42 (d, J = 5.1 Hz, 1H), 7.25 (t, J = 9.1 Hz, 1H), 6.77
(dd, J' = 2.7 Hz, J'' = 6.2 Hz, 1H), 6.70- 6.78 (m, 1H), 4.15 (q, J
= 7.1 Hz, 4H), 1.08 (t, J = 7.1 Hz, [M + H] 422.2 H 6H) 143
##STR00463## Methyl (3-((3- chloro-4- fluorophenyl) amino)furo
[2,3-c]pyridin- 2-yl)carbamate DMSO-d.sub.6: .delta. 10.24 (s, 1H),
8.85 (s, 1H), 8.34 (d, J = 5.1 Hz, 1H), 7.77 (s, 1H), 7.31 (d, J =
5.1 Hz, 1H), 7.19 (t, J = 9.1 Hz, 1H), 6.73-6.71 (m, 1H), 6.62-6.60
(m, 1H), 3.63 (s, 3H) [M + H] 336 G 144 ##STR00464## Methyl (3-
chloro-4- fluorophenyl) (2-((methoxy- carbonyl) amino)furo
[2,3-c]pyridin- 3-yl)carbamate DMSO-d.sub.6: .delta. 8.97 (s, 1H),
8.42 (d, J = 5.0 Hz, 1H), 8.23 (s, 1H), 7.39 (d, J = 4.8 Hz, 1H),
7.24 (t, J = 8.8 Hz, 1H), 6.79- 6.78 (m, 1H), 6.68- 6.66 (m, 1H),
3.69 (s, 6H) [M + H] 394 H 145 ##STR00465## Ethyl (3- chloro-4-
fluorophenyl) (7-cyano-2- ((ethoxy- carbonyl) amino)furo
[2,3-c]pyridin- 3-yl)carbamate DMSO-d.sub.6: .delta. 8.50 (d, J =
5.1 Hz, 1H), 7.63 (d, J = 5.1 Hz, 1H), 7.11 (t, J = 9 Hz, 1H), 6.84
(dd, J' = 6.3 Hz, J'' = 2.8 Hz, 1H), 6.75 (dt, J' = 8.9 Hz, J'' =
6.8 Hz, J''' = 3.7 Hz, 1H), 6.64 (s, 1H), 4.19 (q, J = 7.1 Hz, 4H),
1.15 (t, J = 7.1 Hz, 6H) [M + H] 447 H 146 ##STR00466## Ethyl (3-
chloro-4- fluorophenyl) (7-(3- cyanophenyl)- 2-((ethoxy- carbonyl)
amino)furo [2,3-c]pyridin- 3-yl)carbamate DMSO-d.sub.6: .delta.
8.60- 8.56 (m, 3H), 8.35 (s, 1H), 8.00 (d, J = 7.5 Hz, 1H), 7.83
(t, J = 7.8 Hz, 1H), 7.50 (d, J = 4.6 Hz, 1H), 7.26 (t, J = 9.6 Hz,
1H), 6.83 (bs, 1H), 6.75 (bs, 1H), 4.17 (q, J = 6.7 Hz, 4H), 1.10
(t, J = 6.8 Hz, 6H) [M + H] 523.2 H 147 ##STR00467## N.sup.3-(3-
Chloro-4- fluorophenyl)- 7-(4- fluorophenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.30 (dd, J' 8.2 Hz,
J'' = 5.8 Hz, 2H), 8.15 (d, J = 5.2 Hz, 1H), 7.38 (t, J = 8.8 Hz,
2H), 7.17-7.11 (m, 4H), 6.87 (d, J = 5.1 Hz, 1H), 6.59 (dd, J' 6.2
Hz, J'' = 2.87 Hz, 1H), 6.52-6.50 (m, 1H) [M + H] 371.8 A 148
##STR00468## N.sup.3-(3- Chloro-4- fluorophenyl)- 7-(1-methyl-
1H-pyrazol-4- yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.30 (s, 1H), 8.12 (s, 1H), 8.02 (s, 1H), 7.12 (bs, 2H),
6.90 (bs, 2H) 6.72 (s, 1H), 6.57 (s, 1H), 6.49 (s, 1H), 3.94 (s,
3H) [M + H] 358 A 149 ##STR00469## 4-(2-Amino-3- ((3-chloro-4-
fluorophenyl) amino)furo [2,3-c]pyridin- 7-yl) benzonitrile
DMSO-d.sub.6: .delta. 8.46 (d, J = 8.2 Hz, 2H), 8.21 (d, J = 4.9
Hz, 1H), 8.01 (d, J = 8.2 Hz, 2H), 7.17-7.11 (m, 4H), 6.93 (d, J =
4.9 Hz, 1H), 6.60-6.59 (m, 1H), 6.52-6.50 (m, 1H) [M + H] 379 C 151
##STR00470## 4-(2-Amino-3- ((3-chloro-4- fluorophenyl) amino)furo
[2,3-c]pyridin- 7-yl)benzamide CDCN.sub.3: .delta. 8.42 (d, J = 8.3
Hz, 2H), 8.25 (d, J = 5.0 Hz, 1H), 7.97 (d, J = 8.4 Hz, 2H), 7.03
(t, J = 9.0 Hz, 1H), 6.98 (d, J = 5.0 Hz, 1H), 6.85 (bs, 1H), 6.66
(dd, J' = 6.3 Hz, J'' = 2.8 Hz, 1H), 6.58 (dt, J' 8.8 Hz, J'' = 6.7
Hz, J''' = 3.3 Hz, 1H), 6.05 (bs, 1H), 5.72 (s, 1H), 5.46 (s, 2H),
[M + H] 397 C 152 ##STR00471## Methyl 4-(2- amino-3-((3- chloro-4-
fluorophenyl) amino)furo [2,3-c]pyridin- 7-yl)benzoate
DMSO-d.sub.6: .delta. 8.42 (d, J = 8.4 Hz, 2H), 8.21 (d, J = 5.0
Hz, 1H), 8.10 (d, J = 8.4 Hz, 2H), 7.16-7.08 (m, 4H), 6.91 (d, J =
5.0 Hz, 1H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.53-6.50
(m, 1H), 3.90 (s, 3H) [M + H] 412 C 153 ##STR00472## N.sup.3-(3-
Chloro-4- fluorophenyl)- 7-iodofuro [2,3-c]pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 7.83 (d, J = 5.0 Hz, 1H), 7.21 (s, 2H),
7.15-7.09 (m, 2H), 6.83 (d, J = 5.0 Hz, 1H), 6.57- 6.56 (m, 1H),
6.47- 6.45 (m, 1H) [M + H] 403.8 C 154 ##STR00473## N.sup.3-(3-
Chloro-4- fluorophenyl)- 7-(2- morpholino- pyridin-4- yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.31 (d, J = 4.6 Hz,
1H), 8.18 (d, J = 4.7 Hz, 1H), 7.59 (s, 1H), 7.51 (s, 1H), 7.37
(bs, 2H), 7.20 (s, 1H), 7.14 (t, J = 8.7 Hz, 1H), 6.95 (d, J = 4.3
Hz, 1H), 6.61 (s, 1H), 6.53 (bs, 1H), 3.75 (s, 4H), 3.53 (s, 4H) [M
+ H] 440 A 155 ##STR00474## Benzyl (3-((3- chloro-4- fluorophenyl)
amino)-7- cyanofuro [2,3-c]pyridin- 2-yl)carbamate CD.sub.3CN:
.delta. 8.42 (d, J = 5.1 Hz, 1H), 8.37 (bh, 1H), 7.50 (d, J = 5.1
Hz, 1H), 7.37 (bs, 5H), 7.05 (t, J = 9.0 Hz, 1H), 6.75 (dd, J' =
6.2 Hz, J'' = 2.7 Hz, 1H), 6.64 (dt, J' = 8.8 Hz, J'' = 6.6 Hz,
J''' = 3.2 Hz, 1H), 6.32 (s, 1H), [M - H] 435.2 G 5.18 (s, 2H) 156
##STR00475## Ethyl (3- chloro-4- fluorophenyl) (2-((ethoxy-
carbonyl) amino)-7- iodofuro[2,3- c]pyridin-3- yl)carbamate
CD.sub.3CN: .delta. 8.20 (d, J = 5.2 Hz, 1H), 7.32 (d, J = 5.1 Hz,
1H), 7.10 (t, J = 9.0 Hz, 1H), 6.84 (dd, J' = 6.2 Hz, J'' = 2.8 Hz,
1H), 6.75 (dt, J' = 8.8 Hz, J'' = 6.8 Hz, J''' = 3.1 Hz, 1H), 6.56
(s, 1H), 4.21 (q, J = 7.1 Hz, 4H), 1.17 (t, J = 7.1 Hz, 6H) [M - H]
546.1 H 157 ##STR00476## Ethyl (7-(4- carbamoyl- phenyl)-3-((3-
chloro-4- fluorophenyl) amino)furo[2,3- c]pyridin-2- yl)carbamate
DMSO-d.sub.6: .delta. 10.34 (s, 1H), 8.47 (d, J = 4.7 Hz, 1H), 8.38
(d, J = 7.8 Hz, 2H), 8.10-8.05 (m, 3H), 7.79 (s, 1H), 7.47 (s, 1H),
7.34 (d, J = 4.5 Hz, 1H), 7.19 (t, J = 9.0 Hz, 1H), 6.77 (bs, 1H),
6.66 (bs, 1H), 4.10 (q, J = 6.8 Hz, 2H), 1.16 (t, J = 6.8 Hz, 3H)
[M + H] 469 G 158 ##STR00477## N.sup.3-(3- Chloro-4- fluorophenyl)-
7-(2- methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.65 (s, 1H), 8.29-8.13 (m, 3H), 7.03 (bs, 2H),
6.66 (s, 1H), 6.60 (s, 1H), 5.94 (s, 1H), 5.78 (bs, 2H), 2.66 (s,
3H) [M + H] 369 A 159 ##STR00478## 7-Bromo-N.sup.3- (3-chloro-4-
fluorophenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 7.85 (d, J = 5.1 Hz, 1H), 7.26 (bs, 2H), 7.16 (s, 1H), 7.12
(t, J = 9.1 Hz, 1H), 6.88 (d, J = 5.1 Hz, 1H), 6.58 (dd, J' = 6.3
Hz, J'' = 2.7 Hz, 1H), 6.48 (dt, J' = 8.9 Hz, J'' = 6.8 Hz, J''' =
3.4 Hz, 1H) [M + H] 356 A 160 ##STR00479## Ethyl (7-((4- carbamoyl-
phenyl)ethynyl)- 2-((ethoxy- carbonyl)amino) furo[2,3-c]
pyridin-3-yl) (3-chloro-4- fluorophenyl) carbamate DMSO-d.sub.6:
.delta. 8.47 (d, J = 5.1 Hz, 1H), 8.36 (s, 1H), 8.12 (s, 1H), 7.97
(d, J = 8.0 Hz, 2H), 7.76 (d, J = 8.0 Hz, 2H), 7.52 (s, 1H), 7.49
(d, J = 5.1 Hz, 1H), 7.26 (t, J = 9.0 Hz, 1H), 6.82 (dd, J' = 6.2
Hz, J'' = 2.5 Hz, 1H), 6.75- 6.71 (m, 1H), 4.18 (q, J = 7.0 Hz,
4H), 1.11 (t, J = 7.0 Hz, 6H) [M - H] 563.2 H 161 ##STR00480##
Ethyl (7- bromo-2- ((ethoxy- carbonyl) amino)furo [2,3-c]pyridin-
3-yl)(3-chloro- 4-fluorophenyl) carbamate DMSO-d.sub.6: .delta.
8.38 (s, 1H), 8.24 (d, J = 5.0 Hz, 1H), 7.48 (d, J = 5.2 Hz, 1H),
7.25 (t, J = 9.1 Hz, 1H), 6.82-6.81 (m, 1H), 6.73-6.71 (m, 1H),
4.17 (q, J = 7.0 Hz, 4H), 1.10 (t, J = 7.0 Hz, 6H) [M + H] 502.2 H
162 ##STR00481## Ethyl (3-((3- chloro-4- fluorophenyl) amino)-7-
phenylfuro[2,3- c]pyridin-2- yl)carbamate DMSO-d.sub.6: .delta.
1.04 (s, 1H), 8.44 (d, J = 4.8 Hz, 1H), 8.31 (d, J = 7.6 Hz, 2H),
7.78 (s, 1H), 7.58 (t, J = 7.3 Hz, 2H), 7.52-7.50 (m, 1H), 7.30 (d,
J = 4.9 Hz, 1H), 7.19 (t, J = 8.9 Hz, 1H), 6.67-6.76 (m, 1H),
6.66-6.64 (m, 1H), 4.10 (q, J = 6.9 Hz, 2H), 1.15 (t, J = 6.9 Hz,
3H) [M + H] 426 G 163 ##STR00482## Ethyl (3-((3- chloro-4-
fluorophenyl) amino)-7-(2,6- dimethyl- pyridin-4-yl) furo[2,3-c]
pyridin-2- yl)carbamate CD.sub.3CN: .delta. 8.42 (d, J = 5.0 Hz,
1H), 7.95 (s, 2H), 7.27 (d, J = 5.1 Hz, 1H), 7.06 (t, J = 9.0 Hz,
1H), 6.77 (dd, J' = 6.2 Hz, J'' = 2.9 Hz, 1H), 6.69-6.65 (m, 1H),
6.32 (s, 1H), 4.18 (q, J = 7.0 Hz, 2H), 2.57 (s, 6H), 1.23 (t, J =
7.0 Hz, 3H) [M + H] 455 G 164 ##STR00483## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(2,6- dimethyl- pyridin-4-yl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.19 (d, J = 5.0 Hz,
1H), 7.88 (s, 2H), 7.16-7.12 (m, 4H), 6.92 (d, J = 4.9 Hz, 1H),
6.60 (dd, J' = 6.1 Hz, J'' = 2.4 Hz, 1H), 6.52- 6.49 (m, 1H), 2.53
(s, 6H) [M + H] 383 A 165 ##STR00484## Ethyl (3- chloro-4-
fluorophenyl) (7-(2,6- dimethyl- pyridin-4-yl)- 2-((ethoxy-
carbonyl) amino)furo [2,3-c]pyridin- 3-yl)carbamate CD.sub.3CN:
.delta. 8.50 (d, J = 5.0 Hz, 1H), 7.92 (s, 2H), 7.39 (d, J = 4.9
Hz, 1H), 7.09 (t, J = 9.0 Hz, 1H), 6.85-6.84 (m, 1H), 6.76-6.74 (m,
1H), 6.54 (s, 1H), 4.19 (q, J = 7.1 Hz, 4H), 2.57 (s, 6H), 1.14 (t,
J = 7.1 Hz, 6H) [M + H] 527 H 166 ##STR00485## 7-Chloro-N3-
(3-chloro-4- fluorophenyl) furo[2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 7.87 (d, J = 5.1 Hz, 1H), 7.25 (s, 2H), 7.16-
7.10 (m, 2H), 6.88 (d, J = 5.1 Hz, 1H), 6.59 (dd, J' = 6.2 Hz, J''
= 2.6 Hz, 1H), 6.48 (dt, J' = 8.8 Hz, J'' = 6.7 Hz, J''' = 3.4 Hz,
1H) [M + H] 312 A 167 ##STR00486## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-iodo-5- methoxyfuro [2,3-c]pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 7.23 (s, 2H), 7.14-7.09 (m, 2H), 6.56-6.55
(m, 1H), 6.46-6.43 (m, 1H), 6.03 (s, 1H), 3.74 (s, 3H) [M + H] 434
A
169 ##STR00487## 4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo [2,3-c]pyridin- 7-yl)-N-(2- methoxyethyl) benzamide
CD.sub.3CN: .delta. 8.11 (d, J = 8.2 Hz, 2H), 8.06-8.03 (m, 3H),
7.26 (bs, 1H), 7.16 (d, J = 6.2 Hz, 1H), 7.08 (t, J = 9.0 Hz, 1H),
6.70 (dd, J' = 6.1 Hz, J'' = 2.6 Hz, 1H), 6.65-6.60 (m, 3H), 5.88
(s, 1H), 3.59-3.54 (m, 4H), 3.35 (s, 3H) [M + H] 455 A 170
##STR00488## N.sup.3-(3-chloro-4- fluorophenyl)- 7-(3,4-
difluorophenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.26 (t, J = 10.4 Hz, 1H), 8.17 (d, J = 5.2 Hz, 2H), 7.62
(q, J = 9.3 Hz, 1H), 7.16-7.11 (m, 4H), 6.89 (d, J = 5.0 Hz, 1H),
6.60 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.53-6.50 (m, 1H), [M +
H] 390 A 171 ##STR00489## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(2,3,4- trifluorophenyl) furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.17 (d, J = 5.1 Hz, 1H), 7.63-7.57 (m, 1H),
7.50 (q, J = 8.5 Hz, 1H), 7.15-7.11 (m, 2H), 7.07 (s, 2H), 6.94 (d,
J = 5.0 Hz, 1H), 6.59 (dd, J' = 6.2 Hz, J'' = 2.7 Hz, 1H),
6.51-6.48 (m, 1H) [M + H] 408 A 172 ##STR00490##
7-Bromo-N.sup.3-(3- chloro-4- fluorophenyl)- 5-methoxy- furo[2,3-c]
pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.27 (s, 2H), 7.14-7.08
(m, 2H), 6.57 (dd, J' = 6.3 Hz, J'' = 2.7 Hz, 1H), 6.46 (dt, J' =
8.7 Hz, J'' = 6.6 Hz, J''' = 3.5 Hz, 1H), 6.08 (s, 1H), 3.75 (s,
3H) [M + H] 386 A 173 ##STR00491## 4-(2-Amino-3- ((3-chloro-4-
fluorophenyl) amino)-5- methoxyfuro [2,3-c]pyridin- 7-yl)
benzonitrile DMSO-d.sub.6: .delta. 8.47 (d, J = 8.2 Hz, 2H), 8.01
(d, J = 8.2 Hz, 2H), 7.18-7.09 (m, 4H), 6.61-6.59 (m, 1H),
6.51-6.48 (m, 1H), 6.14 (s, 1H), 3.88 (s, 3H) [M + H] 409 A 174
##STR00492## N.sup.3-(3-chloro-4- fluorophenyl)- 7-(4-
fluorophenyl)- 5-methoxy- furo[2,3-c] pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 8.34 (dd, J' = 8.2 Hz, J'' = 5.6 Hz, 2H),
7.37 (t, J = 8.8 Hz, 2H), 7.15-7.05 (m, 4H), 6.59-6.57 (m, 1H),
6.50-6.48 (m, 1H), 6.07 (s, 1H), 3.87 (s, 3H) [M + H] 402 A 175
##STR00493## 4-(2-amino-3- ((3-chloro-4- fluorophenyl) amino)-5-
methoxyfuro [2,3-c]pyridin- 7-yl)benzamide DMSO-d.sub.6: .delta.
8.36 (d, J = 8.3 Hz, 2H), 8.06 (s, 1H), 8.01 (d, J = 8.3 Hz, 2H),
7.45 (s, 1H), 7.15-7.08 (m, 4H), 6.59 (dd, J' = 6.1 Hz, J'' = 2.6
Hz, 1H), 6.51-6.49 (m, 1H), 6.11 (s, 1H), 3.89 (s, 3H) [M + H] 427
A 176 ##STR00494## N.sup.3-(3-chloro-4- fluorophenyl)- 7-fluorofuro
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.66 (d, J = 5.2
Hz, 1H), 7.15-7.11 (m, 4H), 6.83 (dd, J' = 5.1 Hz, J'' = 2.6 Hz,
1H), 6.58 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.48 (dt, J' = 8.9
Hz, J'' = 6.7 Hz, J'' = 3.4 Hz, 1H) [M + H] 296 C 177 ##STR00495##
5-Chloro-N3- (3-chloro-4- fluorophenyl) furo[2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.20 (s, 1H), 7.27 (s, 2H), 7.13 (t,
J = 9.0 Hz, 1H), 7.08 (s, 1H), 6.77 (s, 1H), 6.58- 6.57 (m, 1H),
6.47- 6.45 (m, 1H) [M + H] 311.8 A 178 ##STR00496## Ethyl (3-
chloro-4- fluorophenyl) (6-cyano-2- ((ethoxy- carbonyl)amino)
benzofuran-3- yl)carbamate DMSO-d.sub.6: .delta. 8.28 (s, 2H), 7.70
(d, J = 7.4 Hz, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.23 (t, J = 9.4 Hz,
1H), 6.78 (bs, 1H), 6.71 (bs, 1H), 4.16-4.14 (m, 4H), 1.09 (bs, 6H)
[M - H] 444 H 179 ##STR00497## N.sup.3-(3-chloro-4- fluorophenyl)-
6-nitrobenzo- furan-2,3- diamine DMSO-d.sub.6: .delta. 8.11 (d, J =
1.9 Hz, 1H), 7.98 (dd, J' = 8.6 Hz, J'' = 1.9 Hz, 1H), 7.35 (bs,
2H), 7.19 (s, 1H), 7.13 (t, J = 9.0 Hz, 1H), 6.92 (d, J = 8.6 Hz,
1H), 6.59 (dd, J' = 6.2 Hz, J'' = 2.7 [M - H] 320 B Hz, 1H), 6.49
(dt, J' = 8.9 Hz, J'' = 6.7 Hz, J''' = 3.5 Hz, 1H), 180
##STR00498## N.sup.3-(3-Chloro-4- fluorophenyl)- 5-fluoro-7-
(pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.76 (d, J = 5.8 Hz, 2H), 8.15 (d, J = 5.8 Hz, 2H), 7.50
(s, 2H), 7.17- 7.12 (m, 2H), 6.63 (dd, J' = 6.1 Hz, J'' = 2.5 Hz,
1H), 6.53- 6.49 (m, 2H) [M + H] 373.2 C 181 ##STR00499##
N.sup.3-(3-Chloro-4- fluorophenyl)- 5-fluorofuro [2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 7.99 (s, 1H), 7.24 (s,
2H), 7.15-7.10 (m, 2H), 6.58-6.57 (m, 1H), 6.47-6.45 (m, 1H), 6.38
(s, 1H) [M + H] 296.2 C 182 ##STR00500## 2-Amino-3-((2-
chlorophenyl) amino)furo [2,3-c]pyridine- 7-carbonitrile
DMSO-d.sub.6: .delta. 8.14 (d, J = 5.1 Hz, 1H), 7.61 (s, 2H), 7.31
(d, J = 7.89 Hz, 1H), 7.05- 7.01 (m, 2H), 6.73 (s, 1H), 6.66 (t, J
= 7.5 Hz, 1H), 6.35 (d, J = 8.1 Hz, 1H) [M + H] 285 A 183
##STR00501## 2-Amino-3- ((4-fluoro-3- (trifluoro- methyl)phenyl)
amino)furo [2,3-c]pyridine- 7-carbonitrile DMSO-d.sub.6: .delta.
8.15 (d, J = 5.1 Hz, 1H), 7.63 (s, 2H), 7.38 (s, 1H), 7.22 (t, J =
9.6 Hz, 1H), 7.11 (d, J = 4.8 Hz, 1H), 6.84 (s, 1H), 6.76-6.73 (m,
1H) [M - H] 335 A 184 ##STR00502## N.sup.3-(3,4- difluoro-
phenyl)-7- (pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.72 (s, 2H), 8.26 (s, 3H), 7.07-7.03 (m, 2H),
6.47-6.40 (m, 2H), 5.74 (s, 1H), 5.55 (s, 2H) [M + H] 339 A 185
##STR00503## 2-Amino-3- ((3,4- difluorophenyl) amino)furo
[2,3-c]pyridine- 7-carbonitrile DMSO-d.sub.6: .delta. 8.14 (d, J =
5.2 Hz, 1H), 7.59 (s, 2H), 7.18-7.10 (m, 2H), 7.07 (d, J = 5.1 Hz,
1H), 6.48- 6.43 (m, 1H), 6.34- 6.32 (m, 1H) [M + H] 287.1 A 186
##STR00504## 2-Amino-3- ((3,5- difluorophenyl) amino)furo
[2,3-c]pyridine- 7-carbonitrile DMSO-d.sub.6: .delta. 8.16 (d, J =
5.1 Hz, 1H), 7.64 (s, 2H), 7.59 (s, 1H), 7.10 (d, J = 5.4 Hz, 1H),
6.37-6.32 (m, 1H), 6.18-6.15 (m, 2H) [M - H] 285.1 A 187
##STR00505## N.sup.3-(3,5- Difluoro- phenyl)-7- (pyridin-4-
yl)furo[2,3- c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.75 (d,
J = 4.3 Hz, 2H), 8.23 (d, J = 3.7 Hz, 3H), 7.60 (s, 1H), 7.22 (s,
2H), 6.96 (d, J = 4.4 Hz, 1H), 6.34 (t, J = 9.4 Hz, 1H), 6.16 (d, J
= 9.3 Hz, 2H) [M + H] 339.1 A 188 ##STR00506## N.sup.3-(3,4-
Difluoro- phenyl)-5- fluoro-7- (pyridin-4- yl)furo[2,3- c]pyridine-
2,3-diamine DMSO-d.sub.6: .delta. 8.76 (d, J = 4.8 Hz, 2H), 8.14
(d, J = 4.9 Hz, 2H), 7.48 (s, 2H), 7.16- 7.11 (m, 2H), 6.51- 6.47
(m, 2H), 6.36- 6.34 (m, 1H) [M + H] 357.2 C 189 ##STR00507##
7-Fluoro-N.sup.3- (4-fluoro-3- (trifluoro- methyl)phenyl)
furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 7.67 (d, J
= 4.9 Hz, 1H), 7.34 (s, 1H), 7.24-7.18 (m, 3H), 6.85-6.82 (m, 2H),
6.74-6.71 (m, 1H) [M - H] 328.2 C 190 ##STR00508##
N.sup.3-(3-Chloro-4- fluorophenyl)- 5,7-difluoro- furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 7.46 (s, 2H), 7.16-7.11
(m, 2H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.7 Hz, 1H), 6.48 (dt, J' =
8.8 Hz, J'' = 6.8 Hz, J''' = 3.4 Hz, 1H), 6.42 (s, 1H) [M + H] 314
C 191 ##STR00509## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(2,6-
difluoro- pyridin-4- yl)furo[2,3- c]pyridine- 2,3-diamine
DMSO-d.sub.6: .delta. 8.23 (d, J = 4.9 Hz, 1H), 7.93 (s, 2H), 7.31
(bs, 2H), 7.19 (s, 1H), 7.14 (t, J = 9.1 Hz, 1H), 7.01 (d, J = 4.9
Hz, 1H), 6.60 (dd, , J' = 6.1 Hz, J'' = 2.3 Hz, 1H), 6.52-6.50 (m,
1H) [M + H] 391 C 192 ##STR00510## N.sup.3-(4-Fluoro-3- (trifluoro-
methyl)phenyl)- 7-(pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.75 (d, J = 5.1 Hz, 2H), 8.23 (bs, 3H), 7.36
(s, 1H), 7.24-7.20 (m, 3H), 6.97 (d, J = 4.7 Hz, 1H), 6.84 (bs,
1H), 6.77-6.75 (m, 1H) [M + H] 389 A 193 ##STR00511## Ethyl (2-
((ethoxy- carbonyl) amino)-7- (pyridin-4- yl)furo[2,3- c]pyridin-3-
yl)(4-fluoro-3- (trifluoro- methyl)phenyl) carbamate DMSO-d.sub.6:
.delta. 8.82 (bs, 2H), 8.60 (d, J = 5.1 Hz, 1H), 8.54 (s, 1H), 8.24
(s, 2H), 7.56 (d, J = 5.1 Hz, 1H), 7.36 (t, J = 9.5 Hz, 1H),
7.05-7.02 (m, 2H), 4.16 (q, J = 7.1 Hz, 4H), 1.07 (t, J = 7.1 Hz,
6H) [M + H] 533.2 H 194 ##STR00512## Ethyl (7-cyano- 2-((ethoxy-
carbonyl) amino)furo [2,3-c]pyridin- 3-yl)(4-fluoro- 3-(trifluoro-
methyl)phenyl) carbamate CD.sub.3CN: .delta. 8.50 (d, J = 5.1 Hz,
1H), 7.62 (d, J = 5.1 Hz, 1H), 7.18 (t, J = 10.0 Hz, 1H), 7.03-
7.01 (m, 2H), 6.76 (s, 1H), 4.19 (q, J = 7.1 Hz, 4H), 1.14 (t, J =
7.1 Hz, 6H) [M - H] 479.2 H 195 ##STR00513## 4-(2-Amino-3-
((3-chloro-4- fluorophenyl) amino)furo[2,3- c]pyridin-7-yl)-
N-methyl- picolinamide DMSO-d.sub.6: .delta. 8.88 (s, 1H),
8.83-8.78 (m, 2H), 8.40 (d, J = 4.4 Hz, 1H), 8.25 (d, J = 4.9 Hz,
1H), 7.18- 7.11 (m, 4H), 6.98 (d, J = 4.9 Hz, 1H), 6.62-6.60 (m,
1H), 6.53-6.51 (m, 1H), 2.88 (d, J = 4.6 Hz, 3H) [M + H] 412 J 196
##STR00514## N.sup.3-(3,4- Difluoro- phenyl)-7-(2- methoxy-
phenyl)furo [2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.09
(d, J = 5.1 Hz, 1H), 7.44 (t, J = 7.7 Hz, 1H), 7.38 (d, J = 7.4 Hz,
1H), 7.17-7.11 (m, 3H), 7.06 (t, J = 7.4 Hz, 1H), 6.84-6.82 (m,
3H), 6.44-6.40 (m, 1H), 6.32-6.30 (m, 1H), 3.76 (s, 3H) [M + H]
368.1 A 197 ##STR00515## 7-(2- Chlorophenyl)- N.sup.3-(3,4-
difluorophenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.13 (d, J = 5.1 Hz, 1H), 7.61-.55 (m, 2H), 7.52-7.45 (m,
2H), 7.18-7.11 (m, 2H), 6.96 (bs, 2H), 6.90 (d, J = 5.0 Hz, 1H),
6.46-6.41 (m, 1H), 6.33-6.31 (m, 1H) [M + H] 372 A 198 ##STR00516##
N.sup.3-(3,4- Difluoro- phenyl)-7-(2- methylpyridin- 4-yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.60 (d, J = 5.0 Hz,
1H), 8.20 (d, J = 5.0 Hz, 1H), 8.09 (s, 1H), 8.03 (d, J = 4.8 Hz,
1H), 7.17-7.10 (m, 4H), 6.93 (d, J = 5.0 Hz, 1H), 6.47-6.42 (m,
1H), 6.34-6.32 (m, 1H), 2.58 (s, 3H) [M + H] 353.1 A 199
##STR00517## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(3- (trifluoro-
methyl)phenyl) furo[2,3-c] pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.60- 8.57 (m, 2H), 8.21 (d, J = 5.0 Hz, 1H), 7.84-7.77 (m,
2H), 7.17-7.12 (m, 4H), 6.92 (d, J = 5.0 Hz, 1H), 6.61 (dd, J' =
6.3 Hz, J'' = 2.6 Hz, 1H), 6.51 (dt, J' = 8.9 Hz, J'' = 6.7 Hz,
J''' = 3.5 Hz, 1H) [M + H] 422 A, C 200 ##STR00518##
N.sup.3-(3-Chloro-4- fluorophenyl)- N7,N7- diphenylfuro
[2,3-c]pyridine- 2,3,7-triamine DMSO-d.sub.6: .delta. 7.82 (d, J =
5.1 Hz, 1H), 7.27 (t, J = 7.8 Hz, 4H), 7.14 (t, J = 9.0 Hz, 1H),
7.05-7.01 (m, 3H), 6.93 (d, J = 7.8 Hz, 4H), 6.72 (d, J = 5.0 Hz,
1H), 6.67 (bs, 2H), 6.59 (dd, J' = 6.3 Hz, J'' = 2.5 Hz, 1H),
6.49-6.44 (m, 1H) [M + H] 445 A 201 ##STR00519## N.sup.3-(4-
Fluorophenyl)- 7-(pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.73 (d, J = 5.3 Hz, 2H), 8.27-8.24 (m, 3H),
7.01 (d, J = 4.8 Hz, 1H), 6.90 (t, J = 8.7 Hz, 2H), 6.62-6.59 (m,
2H), 5.58 (s, 1H), 5.49 (bs, 2H) [M + H] 321.2 A 202 ##STR00520##
N.sup.3-(2- Chlorophenyl)- 7-(pyridin-4- yl)furo[2,3-
c]pyridine-2,3- diamine CD.sub.3CN: .delta. 8.78 (d, J = 6.2 Hz,
2H), 8.29-8.25 (m, 3H), 7.35 (d, J = 9.0 Hz, 1H), 7.07-7.04 (m,
2H), 6.72 (t, J = 6.7 Hz, 1H), 6.49 (d, J = 7.0 Hz, 1H), 5.89 (s,
1H), 5.83 (bs, 2H), [M + H] 337 A 203 ##STR00521## 7-(Pyridin-4-
yl)-N.sup.3-(3- (trifluoro- methyl)phenyl) furo[2,3-c]
pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.75 (d, J = 4.9 Hz,
2H), 8.24-8.21 (m, 3H), 7.44 (s, 1H), 7.30 (t, J = 7.8 Hz, 1H),
7.19 (bs, 2H), 6.96-6.90 (m, 2H), 6.84 (s, 1H), 6.77 (d, J = 7.5
Hz, 1H) [M + H] 371 A 204 ##STR00522## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(naphthalen- 1-yl)furo[2,3- c]pyridine-2,3-
diamine CD.sub.3CN: .delta. 8.30 (d, J = 5.1 Hz, 1H), 8.04-7.99 (m,
2H), 7.92 (d, J = 8.4 Hz, 1H), 7.74 (d, J = 6.8 Hz, 1H), 7.64 (t, J
= 7.7 Hz, 1H), 7.56 (t, J = 7.5 Hz, 1H), 7.48 (t, J = 7.3 Hz, 1H),
7.07-7.03 (m, 2H), 6.71 (dd, J' = 6.1 Hz, J'' = 2.6 Hz, 1H),
6.64-6.61 (m, 1H), 5.75 (s, 1H), 5.26 (s, 2H) [M + H] 404 A 205
##STR00523## N.sup.3-(4-fluoro- 3-(trifluoro- methyl)phenyl)-
7-(2-methyl- pyridin-4-yl) furo[2,3-c] pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.73 (d, J = 5.7 Hz, 1H), 8.34 (s, 1H), 8.31-
8.27 (m, 2H), 7.13- 7.09 (m, 2H), 6.87- 6.85 (m, 2H), 6.00 (bs,
3H), 2.75 (s, 3H) [M + H] 403 A 206 ##STR00524## N.sup.3-(2-
chlorophenyl)- 7-(2-methyl- pyridin-4-yl) furo[2,3-c] pyridine-2,3-
diamine CD.sub.3CN: .delta. 8.75 (d, J = 5.4 Hz, 1H), 8.40-8.28 (m,
3H), 7.36 (d, J = 7.9 Hz, 1H), 7.14 (d, J = 5.0 Hz, 1H), 7.06 (t, J
= 7.3 Hz, 1H), 6.73 (t, J = 7.3 Hz, 1H), 6.48 (d, J = 8.0 Hz, 1H),
6.09 (bs, 2H), 5.93 (s, 1H), 2.78 (s, 3H) [M + H] 351 A 207
##STR00525## 7-(2- methylpyridin- 4-yl)-N.sup.3-(3- (trifluoro-
methyl)phenyl) furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6:
.delta. 8.76 (d, J = 5.2 Hz, 1H), 8.22 (d, J = 5.6 Hz, 1H),
8.10-7.95 (m, 4H), 7.58 (s, 1H), 7.33 (t, J = 7.7 Hz, 1H), 7.08 (d,
J = 5.4 Hz, 1H), 6.96 (d, J = 7.4 Hz, 1H), 6.87 (s, 1H), 6.81 (d, J
= 8.0 Hz, 1H), 2.66 (s, 3H) [M + H] 385.1 A 208 ##STR00526##
N.sup.3-(4- fluorophenyl)- 7-(2- methylpyridin- 4-yl)furo[2,3-
c]pyridine-2,3- diamine CD.sub.3CN: .delta. 8.74 (d, J = 5.7 Hz,
1H), 8.34 (s, 1H), 8.30 (d, J = 5.6 Hz, 1H), 8.26 (d, J = 5.6 Hz,
1H), 7.12 (d, J = 5.4 Hz, 1H), 6.92 (t, J = 8.8 Hz, 2H), 6.63 (dd,
J' = 8.9 Hz, J'' = 4.5 Hz, 2H), 6.09 (bs, 2H), 5.71 (bs, 1H), 2.76
(s, 3H) [M + H] 335.2 A 209 ##STR00527## Ethyl (3-((3- chloro-4-
fluorophenyl) amino)benzo [b]thiophen-2- yl)carbamate (500 MHz,
DMSO- d.sub.6: .delta. 10.21 (s, 1H), 7.81 (d, J = 7.6 Hz, 1H),
7.54 (s, 1H), 7.27-7.21 (m, 3H), 7.10 (t, J = 9.0 Hz, 1H),
6.51-6.50 (m, 1H), 6.39-6.37 (m, 1H), 4.13 (q, J = 7.0 Hz, 2H),
1.19 (t, J = [M + H] 365.05 G 7.0 Hz, 3H) 210 ##STR00528##
7-Bromo-N.sup.3- (3-chloro-4- fluorophenyl) benzo[b] thiophene-
2,3-diamine hydrochloride (500 MHz, DMSO- d.sub.6: .delta. 7.32 (s,
1H), 7.16 (d, J = 7.8 Hz, 1H), 7.09-7.06 (m, 2H), 6.95 (d, J =
7.5 Hz, 1H), 6.47-6.46 (m, 1H), 6.42-6.39 (m, 1H), 6.16 (s, 2H) [M
+ H] 370.9 E, I 211 ##STR00529## N.sup.3-(3,4- Difluorophenyl)
benzo[b] thiophene-2,3- diamine (500 MHz, CDCl.sub.3: .delta. 7.59
(d, J = 7.4 Hz, 1H), 7.25-7.12 (m, 3H), 6.96-6.91 (m, 1H),
6.41-6.37 (m, 1H), 6.34-6.31 (m, 1H), 4.95 (s, 1H), 4.15 (s, 2H) [M
- H] 275.04 E 212 ##STR00530## 4-(2-Amino-3- ((3-chloro-4-
fluorophenyl) amino)benzo [b]thiophen-7- yl)benzonitrile
DMSO-d.sub.6: .delta. 7.96 (d, J = 8.2 Hz, 2H), 7.84 (d, J = 8.7
Hz, 2H), 7.31 (s, 1H), 7.26 (t, J = 7.6 Hz, 1H), 7.11-7.06 (m, 2H),
7.02 (dd, J' = 7.8 Hz, J'' = 0.9 Hz, 1H), 6.49 (dd, J' = 6.0 Hz,
J'' = 2.7 Hz, 1H), 6.43 (dt, J' = 8.7 Hz, J'' = 6.8 Hz, J''' = 3.7
Hz, 1H), 6.00 (s, 2H) [M - H] 392.04 I 213 ##STR00531##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(pyridin-4- yl)benzo[b]
thiophene-2,3- diamine DMSO-d.sub.6: .delta. 8.68 (d, J = 5.9 Hz,
2H), 7.65 (d, J = 5.9 Hz, 2H), 7.32 (s, 1H), 7.27 (t, J = 7.6 Hz,
1H), 7.12-7.08 (m, 2H), 7.03 (d, J = 7.8 Hz, 1H), 6.50 (dd, J' =
6.0 Hz, J'' = 2.3 Hz, 1H), 6.45-6.42 (m, 1H), 6.01 (s, 2H) [M + H]
370.05 I 214 ##STR00532## N.sup.3-(3-Chloro-4- fluorophenyl)-
5-methylbenzo [b]thiophene- 2,3-diamine DMSO-d.sub.6: .delta. 7.42
(d, J = 8.7 Hz, 1H), 7.20 (s, 1H), 7.07 (t, J = 8.9 Hz, 1H),
6.79-6.77 (m, 2H), 6.45 (dd, J' = 6.4 Hz, J'' = 2.7 Hz, 1H),
6.42-6.38 (m, 1H), 5.80 (s, 2H), 2.22 (s, [M + H] 307.04 E 3H) 215
##STR00533## N.sup.3-(3,4- difluoro- phenyl)-5- methylbenzo[b]
thiophene-2,3- diamine DMSO-d.sub.6: .delta. 7.42 (d, J = 8.7 Hz,
1H), 7.21 (s, 1H), 7.10- 7.03 (m, 1H), 6.79- 6.76 (m, 2H), 6.32-
6.27 (m, 1H), 6.24- 6.21 (m, 1H), 5.78 (s, 2H) , 2.22 (s, 3H) [M +
H] 291.07 E 216 ##STR00534## N.sup.3-(3,4- difluoro- phenyl)-5-
fluorofuro [2,3-c] pyridine-2,3- diamine DMSO-d.sub.6: .delta. 7.99
(s, 1H), 7.23 9s, 2H), 7.17-7.10 (m, 2H), 6.45-6.39 (m, 1H), 6.37
(s, 1H), 6.30- 6.28 (m, 1H) [M + H] 280.2 C 217 ##STR00535##
N.sup.3-(3- Chlorophenyl)- 7-(pyridin-4- yl)furo[2,3-
c]pyridine-2,3- diamine CD.sub.3CN: .delta. 8.73 (d, J = 5.9 Hz,
2H), 8.27-8.25 (m, 3H), 7.12 (t, J = 8.3 Hz, 1H), 7.04 (d, J = 5.0
Hz, 1H), 6.70 (d, J = 7.8 Hz, 1H), 6.60-6.58 (m, 2H), 5.82 (s, 1H),
5.56 (bs, 2H), [M + H] 337 A 218 ##STR00536## N.sup.3-(2-
Chlorophenyl)- 5-fluorofuro [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.00 (s, 1H), 7.30 (d, J = 7.8 Hz, 1H), 7.23
(s, 2H), 7.04 (t, J = 7.5 Hz, 1H), 6.67-6.62 (m, 2H), 6.34-6.32 (m,
2H), [M + H] 278 C 219 ##STR00537## 4-(2-Amino-3- ((2-
chlorophenyl) amino)furo[2,3- c]pyridin-7-yl)- N-methyl-
picolinamide DMSO-d.sub.6: .delta. 8.88 (s, 1H), 8.85 (d, J = 4.8
Hz, 1H), 8.79 (d, J = 5.1 Hz, 1H), 8.41- 8.39 (m, 1H), 8.24 (d, J =
5.0 1H), 7.32 (d, J = 7.7 Hz, 1H), 7.22 (bs, 2H), 7.03 (t, J = 7.7
Hz, 1H), 6.93 (d, J = 5.0 Hz, 1H), 6.71 (s, 1H), 6.65 (t, J = 7.5
Hz, 1H), 6.38 (d, J = 7.8 Hz, 1H), 2.88 (d, J = 4.8 Hz, 3H) [M + H]
394.2 J 220 ##STR00538## N.sup.3-(3- Chlorophenyl)- 5-fluorofuro
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.00 (s, 1H),
7.24 (s, 2H), 7.20 (s, 1H), 7.09 (t, J = 8.0 Hz, 1H), 6.62 (d, J =
7.4 Hz, 1H), 6.49-6.46 (m, 2H), 6.37 (s, 1H), [M + H] 278.1 C 221
##STR00539## 5-Fluoro-N.sup.3-(3- (trifluoro- methyl)phenyl)
furo[2,3-c] pyridine- 2,3-diamine CD.sub.3CN: .delta. 7.97 (s, 1H),
7.30 (t, J = 7.8 Hz, 1H), 6.99 (d, J = 7.6 Hz, 1H), 6.83 (d, J =
7.8 Hz, 2H), 6.47 (s, 1H), 5.89 (s, 1H), 5.61 (s, 2H) [M + H] 311.7
C 222 ##STR00540## 5-Fluoro-N-(4- fluoro-3- (trifluoro-
methyl)phenyl) furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6:
.delta. 8.00 (s, 1H), 7.29-7.19 (m, 4H), 6.81-6.80 (m, 1H),
6.72-6.70 (m, 1H), 6.42 (s, 1H) [M + H] 330 C 223 ##STR00541##
4-(2-Amino-3- ((3,4- difluorophenyl) amino)furo[2,3-
c]pyridin-7-yl)- N-methyl- picolinamide DMSO-d.sub.6: .delta. 8.86
(s, 2H), 8.80 (d, J = 4.8 Hz, 1H), 8.38 (d, J = 3.9 Hz, 1H), 8.24
(d, J = 4.6 Hz, 1H), 7.27-7.13 (m, 4H), 6.97 (d, J = 4.6 Hz, 1H),
6.46 (dd, J' = 12.4 Hz, J'' = 6.2 Hz, 1H), 6.35 (d, J = 7.6 Hz,
1H), 2.87 (d, J = 4.2 Hz, 3H) [M + H] 396.2 J 224 ##STR00542##
4-(2-Amino-3- ((4- fluorophenyl) amino)furo[2,3- c]pyridin-7-yl)-
N-methyl- picolinamide DMSO-d.sub.6: .delta. 8.86 (s, 2H), 8.80 (d,
J = 4.9 Hz, 1H), 8.39 (d, J = 4.0 Hz, 1H), 8.23 (d, J = 5.0 Hz,
1H), 7.19 (bs, 1H), 6.95-6.92 (m, 4H), 6.56-6.53 (m, 2H), 2.87 (d,
J = 4.5 Hz, 3H) [M + H] 378 J 225 ##STR00543## 4-(2-Amino-3- ((3-
chlorophenyl) amino)furo[2,3- c]pyridin-7-yl)- N- methyl-
picolinamide DMSO-d.sub.6: .delta. 8.93 (s, 2H), 8.63 (s, 1H), 8.35
(bs, 2H), 8.24- 8.20 (m, 2H), 7.46 (s, 1H), 7.16-7.09 (m, 2H), 6.69
(d, J = 7.0 Hz, 1H), 6.59-6.57 (m, 2H), 2.88 (d, J = 3.8 Hz, 3H) [M
+ H] 394.2 J 226 ##STR00544## 4-(2-Amino-3- ((3- (trifluoro-
methyl)phenyl) amino)furo[2,3- c]pyridin-7-yl)- N-methyl-
picolinamide DMSO-d.sub.6: .delta. 8.91 (d, J 2H), 8.76 (s, 1H),
8.30 (d, J = 4.2 Hz, 1H), 8.23 (d, J = 5.5 Hz, 1H), 7.95 (bh, 2H),
7.57 (s, 1H), 7.33 (t, J = 7.9 Hz, 1H), 7.07 (d, J = 5.4 Hz, 1H),
6.95 (d, J = 7.6 Hz, 1H), 6.88 (s, 1H, 6.81 (d, J = 8.2 Hz, 1H),
2.88 (d, J = 4.5 Hz, 3H) [M + H] 428 J 227 ##STR00545##
4-(2-amino-3- ((4-fluoro-3- (trifluoro- methyl)phenyl) amino)furo
[2,3-c]pyridin- 7-yl)-N- methyl- picolinamide DMSO-d.sub.6: .delta.
8.96 (s, 2H), 8.63 (bs, 3H), 8.19 (s, 2H), 7.60 (s, 1H), 7.27 (t, J
= 8.8 Hz, 1H), 7.17 (d, J = 4.4 Hz, 1H), 6.91 (s, 1H), 6.86 (s,
1H), 2.88 (s, 3H) [M + H] 445.9 J 228 ##STR00546##
5-Fluoro-N.sup.3-(4- fluorophenyl) furo[2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 7.98 (s, 1H), 7.17 (s, 2H), 6.93 (t,
J = 8.5 Hz, 2H), 6.82 (s, 1H), 6.51-6.49 (m, 2H), 6.31 (s, 1H) [M +
H] 262.1 C 229 ##STR00547## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(2- methoxy- pyridin-4-yl) furo[2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.32 (d, J = 5.4 Hz, 1H), 8.20 (d, J = 5.0
Hz, 1H), 7.85 (d, J = 5.3 Hz, 1H), 7.66 (s, 1H), 7.15-7.11 (m, 4H),
6.93 (d, J = 5.0 Hz, 1H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.5 Hz, 1H),
6.52-6.49 (m, 1H), 3.93 (s, 3H) [M - H] 385.2 A 230 ##STR00548##
N.sup.3-(3- Bromophenyl) benzo[b] thiophene-2,3- diamine (500 MHz,
DMSO- d.sub.6): .delta. 7.56 (d, J = 7.7 Hz, 1H), 7.31 (s, 1H),
7.10 (t, J = 7.0 Hz, 1H), 6.98-6.93 (m, 3H), 6.67 (d, J = 7.7 Hz,
1H), 6.56 (s, 1H), 6.47 (d, J = 8.0 Hz, 1H), 5.84 (s, 2H) [M - H]
316.97 E 231 ##STR00549## N.sup.3-(4- Fluorophenyl) benzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.55 (d, J
= 7.7 Hz, 1H), 7.08 (t, J = 7.2 Hz, 1H), 6.96- 6.94 (m, 3H), 6.86
(t, 8.2 Hz, 2H), 6.44 (bs, 2H), 5.74 (s, 2H), [M + H] 259.07 E 232
##STR00550## N.sup.3-(2- Ethylphenyl) benzo[b] thiophene-
2,3-diamine hydrochloride (500 MHz, DMSO- d.sub.6): .delta. 7.58
(d, J = 8.1 Hz, 1H), 7.06 (t, J = 7.9 Hz, 1H), 7.00- 6.94 (m, 2H),
6.87 (d, J = 7.8 Hz, 1H), 6.78 (d, J = 7.7 Hz, 1H), 6.53 (t, J =
7.3 Hz, 1H), 6.04 (d, J = 7.4 Hz, 1H), 2.69 (q, [M + H] 269.11 E J
= 7.5 Hz, 2H), 1.24 (t, J = 7.5 Hz, 3H) 233 ##STR00551##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-phenylbenzo [b]thiophene-
2,3-diamine (500 MHz, DMSO- d.sub.6): .delta. 7.63 (d, J = 7.4 Hz,
2H), 7.49 (t, J = 7.5 Hz, 2H), 7.41 (t, J = 7.5 Hz, 1H), 7.30 (s,
1H), 7.22 (t, J = 7.6 Hz, 1H), 7.09 (t, J = 9.0 Hz, 1H), 7.01 (d, J
= 7.3 Hz, 1H), 6.96 (d, J = 7.6 Hz, 1H), 6.52-6.50 (m, 1H),
6.46-6.41 (m, 1H), 5.91 (s, 2H) [M + H] 369.06 I, E 234
##STR00552## N.sup.3-(3- Chlorophenyl) benzo[b] thiophene-2,3-
diamine (400 MHz, DMSO- d.sub.6): .delta. 7.58 (dd, J' = 7.8 Hz,
J'' = 0.8 Hz, 1H), 7.35 (s, 1H), 7.12 (t, J = 7.6 Hz, 1H), 7.05 (t,
J = 7.9 Hz, 1H), 7.00-6.95 (m, 2H), 6.58-6.56 (m, 1H), 6.47-6.45 [M
+ H] 275.04 E (m, 1H), 6.42-6.43 (m, 1H), 5.88 (s, 2H) 235
##STR00553## N.sup.3-(3- Methoxy- phenyl)benzo [b]thiophene-
2,3-diamine (500 MHz, DMSO- d.sub.6): .delta. 7.55 (d, J = 8.1 Hz,
1H), 7.08 (t, J = 7.5 Hz, 1H), 7.01 (s, 1H), 6.96-6.90 (m, 3H),
6.14 (d, J = 7.8 Hz, 1H), 6.09 (d, J = 8.0 Hz, 1H), 6.03 (s, 1H),
5.70 (s, 2H), [M + H] 271.09 E 3.58 (s, 3H) 236 ##STR00554##
N.sup.3-(benzo [d][1,3] dioxol-5- yl)benzo[b] thiophene-2,3-
diamine (500 MHz, DMSO- d.sub.6): .delta. 7.54 (d, J = 7.6 Hz, 1H),
7.08 (t, J = 7.2 Hz, 1H), 6.96- 6.92 (m, 2H), 6.79 (s, 1H), 6.59
(d, J = 8.2 Hz, 1H), 6.10 (d, J = 2.5 Hz, 1H), 5.90 (dd, J' = 8.4
Hz, J'' = 2.2 Hz, 1H), 5.78 (s, [M + H] 285.06 E 2H), 5.70 (s, 2H)
237 ##STR00555## N.sup.3-(4-Bromo-2- methylphenyl) benzo[b]
thiophene-2,3- diamine (500 MHz, DMSO- d.sub.6): .delta. 7.56 (d, J
= 7.7 Hz, 1H), 7.15 (s, 1H), 7.07 (t, J = 7.5 Hz, 1H), 6.94-6.93
(m, 2H), 6.85 (d, J = 7.7 Hz, 1H), 6.44 (s, 1H), 5.94 (d, J = 8.6
Hz, 1H), 5.82 (s, 2H), 2.27 (s, 3H) [M + H] 333.01 E 238
##STR00556## N.sup.3-(3- Chlorophenyl)- 7-(2- methylpyridin-
4-yl)furo[2,3- c]pyridine-2,3- diamine CD.sub.3CN: .delta. 8.72 (d,
J = 5.6 Hz, 1H), 8.33 (s, 1H), 8.28 (d, J = 5.4 Hz, 2H), 7.15-7.10
(m, 2H), 6.72 (d, J = 8.8 Hz, 1H), 6.60 (d, J = 6.7 Hz, 2H),
6.00-5.97 (m, 3H), 2.74 (s, 3H) [M + H] 351 A 239 ##STR00557##
7-(2- Methylpyridin- 4-yl)-N.sup.3- phenylfuro[2,3- c]pyridine-2,3-
diamine CD.sub.3CN: .delta. 8.73 (d, J = 5.7 Hz, 1H), 8.32 (s, 1H),
8.28-8.25 (m, 2H), 7.15 (t, J = 7.8 Hz, 2H), 7.11 (d, J = 5.5 Hz,
1H), 6.73 (t, J = 7.3 Hz, 1H), 6.64 (d, J = 8.0 Hz, 2H), 6.05 (s,
2H), 5.75 (s, 1H), 2.75 (s, 3H) [M + H] 317.2 A 240 ##STR00558##
N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(2- methylpyridin-
4-yl)benzo[b] thiophene-2,3- diamine DMSO-d.sub.6: .delta. 8.53 (d,
J = 5.3 Hz, 1H), 7.52 (s, 1H), 7.44 (d, J = 4.4 Hz, 1H), 7.30 (s,
1H), 7.25 (t, J = 7.7 Hz, 1H), 7.11-7.06 (m, 2H), 7.01 (d, J = 7.6
Hz, 1H), 6.49 (dd, J' = 6.3 Hz, J'' = 2.6 Hz, 1H), 6.43 (dt, J' =
8.8 Hz, J'' = 6.0 Hz, J''' = 3.4 Hz, 1H), 5.99 (s, 2H), 2.51 (s,
3H) [M + H] 384.07 I 241 ##STR00559## 5-Fluoro-N.sup.3-(4-
fluorophenyl)- 7-(2- methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.61 (d, J = 5.2 Hz, 1H), 8.00 (s,
1H), 7.96 (d, J = 5.2 Hz, 1H), 7.41 (s, 2H), 6.94 (t, J = 8.7 Hz,
2H), 6.89 (s, 1H), 6.56-6.63 (m, 2H), 6.40 (d, J = 2.2 Hz, 1H),
2.58 (s, 3H) [M + H] 353.1 C 242 ##STR00560## 7-(2- Methylpyridin-
4-yl)-N.sup.3-(2- (piperidin-1- yl)phenyl)furo [2,3-c]pyridine-
2,3-diamine CD.sub.3CN: .delta. 8.60 (d, J = 5.1 Hz, 1H), 8.22 (d,
J = 5.0 Hz, 1H), 8.15 (s, 1H), 8.08 (d, J = 5.3 Hz, 1H), 7.11 (d, J
= 7.8 Hz, 1H), 6.92 (d, J = 5.0 Hz, 1H), 6.84 (t, J = 6.9 Hz, 1H),
6.69 (t, J = 7.1 Hz, 1H), 6.36 (d, J = 7.8 Hz, 1H), 6.03 (s, 1H),
5.46 (d, J = 2H), 2.93 (bs, 4H), 2.62 (s, 3H), 1.78- 1.74 (m, 4H),
1.62- 1.59 (m, 2H) [M + H] 399.9 A 243 ##STR00561## N.sup.3-(3-
Fluorophenyl)- 7-(2- methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3-
diamine CD.sub.3CN: .delta. 8.74 (d, J = 5.6 Hz, 1H), 8.35 (s, 1H),
8.31-8.27 (m, 2H), 7.17-7.12 (m, 2H), 6.51-6.43 (m, 2H), 6.35 (dt,
J' = 12.0 Hz, J'' = 4.4 Hz, J''' = 2.4 Hz, 1H), 6.06 (bs, 2H), 5.99
(s, 1H), 2.75 (s, 3H) [M + H] 335.1 A 244 ##STR00562##
N.sup.3-(3-Chloro-4- fluorophenyl)- 5-fluoro-7-(2- methylpyridin-
4-yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.84
(d, J = 5.2 Hz, 1H), 8.39 (s, 1H), 8.32 (bs, 1H), 7.64 (s, 2H),
7.20 (s, 1H), 7.15 (t, 9.1 Hz, 1H), 6.62 (dd, J' = 6.2 Hz, J'' =
2.6 Hz, 1H), 6.60 (s, 1H), 6.56-6.52 (m, 1H), 2.74 (s, 3H) [M + H]
387.1 C 245 ##STR00563## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-morpholino- furo[2,3-c] pyridine- 2,3-diamine DMSO-d.sub.6:
.delta. 7.68 (d, J = 5.2 Hz, 1H), 7.11 (t, J = 9.0 Hz, 1H), 7.05
(s, 1H), 6.59 (s, 2H), 6.53 (dd, J' = 6.0 Hz, J'' = 2.0 Hz, 1H),
6.48-6.45 (m, 1H), 6.41 (d, J = 4.8 Hz, 1H), 3.75 (d, J = 4.4 Hz,
4H), 3.61 (d, J = 4.4 Hz, 4H) [M + H] 363.2 A 246 ##STR00564##
N.sup.3-(3,4- Difluoro- phenyl)-5- fluoro-7-(2- methylpyridin-
4-yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.61
(d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.96 (d, J = 4.8 Hz, 1H), 7.47
(s, 2H), 7.17-7.11 (m, 2H), 6.50-6.46 (m, 2H), 6.36-6.33 (m, 1H),
2.59 (s, 3H) [M + H] 371.2 C 247 ##STR00565## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(naphthalen- 2-yl)furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.84 (s, 1H), 8.44 (d, J = 8.5 Hz,
1H), 8.23 (d, J = 5.0 Hz, 1H), 8.07- 8.05 (m, 2H), 8.00- 7.97 (m,
1H), 7.60- 7.57 (m, 2H), 7.19 (s, 1H), 7.15 (t, J = 9.1 Hz, 1H),
7.08 (bs, 2H), 6.90 (d, J = 4.9 Hz, 1H), 6.63-61 (m, 1H), 6.54-6.52
(m, 1H) [M + H] 404 A 248 ##STR00566## N.sup.3-(3- Chlorophenyl)-
7-(2- methylpyridin- 4-yl)benzo[b] thiophene-2,3- diamine
DMSO-d.sub.6: .delta. 8.53 (d, J = 5.3 Hz, 1H), 7.52 (s, 1H), 7.45
(d, J = 4.8 Hz, 1H), 7.40 (s, 1H), 7.25 (t, J = 7.7 Hz, 1H),
7.10-7.06 (m, 1H), 7.04-6.99 (m, 2H), 6.55 (d, J = 7.8 Hz, 1H),
6.45 (d, J = 8.0 Hz, 1H), 6.42 (s, 1H), 5.98 (s, 2H), 2.52 (s, 3H)
[M + H] 366.08 A 249 ##STR00567## 4-(2-Amino-3- ((3-chloro-4-
fluorophenyl) amino)furo [2,3-c]pyridin- 7-yl)-N- phenyl-
picolinamide DMSO-d.sub.6: .delta. 10.72 (s, 1H), 9.01 (s, 1H),
8.91 (d, J = 5.2 Hz, 1H), 8.48 (dd, J' = 5.1 Hz, J'' = 1.5 Hz, 1H),
8.27 (d, J = 5.0 Hz, 1H), 7.95 (d, J = 7.7 Hz, 2H), 7.39 (t, J =
8.0 Hz, 2H), 7.25 (bs, 2H), 7.20 (s, 1H), 7.17-7.12 (m, 2H), 7.00
(d, J = 5.0 Hz, 1H), 6.62 (dd, J' = 6.3 Hz, J'' = 2.7 Hz, 1H),
6.55-6.52 (m, 1H) [M + H] 474.2 J 250 ##STR00568## 4-(2-Amino-3-
((3-chloro-4- fluorophenyl) amino)furo[2,3- c]pyridin-7-yl)-
N-butyl- picolinamide DMSO-d.sub.6:
.delta. 8.87- 8.83 (m, 2H), 8.80 (d, J = 5.0 Hz, 1H), 8.39 (d, J =
4.2 Hz, 1H), 8.25 (d, J = 5.0 Hz, 1H), 7.26 (bs, 2H), 7.19 (s, 1H),
7.14 (t, J = 9.1 Hz, 1H), 6.98 (d, J = 4.9 Hz, 1H), 6.61 (dd, J' =
6.6 Hz, J'' = 2.8 Hz, 1H), 6.53-6.51 (m, 1H), 3.37-3.32 (m, 2H,
merged with DMSO water), 1.57- 1.52 (m, 2H), 1.36- 1.31 (m, 2H),
0.91 (t, [M + H] 454.4 J J = 7.3 Hz, 3H) 251 ##STR00569##
4-(2-Amino-3- ((3-chloro-4- fluorophenyl) amino)furo[2,3-
c]pyridin-7-yl)- N-(tert-butyl) picolinamide DMSO-d.sub.6: .delta.
8.82 (d, J = 4.4 Hz, 2H), 8.35 (d, J = 4.2 Hz, 1H), 8.23 (d, J =
5.4 Hz, 1H), 8.15 (s, 1H), 7.62 (bs, 2H), 7.25 (s, 1H), 7.15 (t, J
= 9.1 Hz, 1H), 7.02 (d, J = 5.3 Hz, 1H), 6.63 (dd, J' = 6.2 Hz, J''
= 2.5 Hz, 1H), 6.56- 6.53 (m, 1H), 1.45 (s, 9H) [M + H] 454.3 J 252
##STR00570## 4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo[2,3- c]pyridin-7-yl)- N-isobutyl- picolinamide
DMSO-d.sub.6: .delta. 8.89 (s, 1H), 8.85-8.79 (m, 2H), 8.41 (d, J =
3.8 Hz, 1H), 8.25 (d, J = 4.9 Hz, 1H), 7.20- 7.18 (m, 3H), 7.14 (t,
J = 9.1 Hz, 1H), 6.98 (d, J = 4.9 Hz, 1H), 6.62-6.61 (m, 1H),
6.53-6.51 (m, 1H), 3.18 (t, J = 6.3 Hz, 2H), 1.95-1.88 (m, 1H),
0.91 (d, J = 6.6 Hz, 3H) [M + H] 454.3 J 253 ##STR00571##
4-(2-Amino-3- ((3-chloro-4- fluorophenyl) amino)furo[2,3-
c]pyridin-7-yl)- N-propyl- picolinamide CD.sub.3CN: .delta.
8.83-8.81 (m, 2H), 8.26 (s, 1H), 8.21 (dd, J' = 5.0 Hz, J'' = 1.7
Hz, 1H), 8.14 (d, J = 5.9 Hz, 1H), 7.15 (d, J = 5.8 Hz, 1H), 7.06
(t, J = 9.0 Hz, 1H), 6.68 (dd, J' = 6.2 Hz, J'' = 2.8 Hz, 1H),
6.63- 6.60 (m, 1H), 6.43 (s, 2H), 5.83 (s, 1H), 3.40 (q, J = 6.8
Hz, 2H), 1.64 (sextet, J = 7.3 Hz, 2H), 0.96 (t, J = 7.4 Hz, 3H) [M
+ H] 440.3 J 254 ##STR00572## N.sup.3-(3- Ahlorophenyl)-
5-fluoro-7-(2- methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.62 (d, J = 5.2 Hz, 1H), 8.00 (s,
1H), 7.96 (d, J = 5.0 Hz, 1H), 7.49 (s, 2H), 7.27 (s, 1H), 7.11 (t,
J = 8.0 Hz, 1H), 6.64 (d, J = 1.2 Hz, 1H), 6.55-6.50 (m, 2H), 6.47
(s, 1H), 2.59 (s, 3H) [M + H] 369 C 256 ##STR00573## N.sup.3-(2,4-
Difluoro- phenyl)-7-(2- methylpyridin- 4-yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.60 (d, J = 4.8 Hz,
1H), 8.19 (d, J = 4.8 Hz, 1H), 8.09 (s, 1H), 8.03 (d, J = 5.2 Hz,
1H), 7.18-7.11 (m, 3H), 6.93 (d, J = 4.8 Hz, 1H), 6.79-6.75 (m,
2H), 6.37-6.31 (m, 1H), 2.59 (s, 3H) [M + H] 353 A 257 ##STR00574##
4-(2-Amino-3- ((3-chloro-4- fluorophenyl) amino)furo[2,3-
c]pyridin-7-yl)- N-cyclohexyl- picolinamide DMSO-d.sub.6: .delta.
8.88 (s, 1H), 8.79 (d, J = 5.1 Hz, 1H), 8.52 (d, J = 8.6 Hz, 1H),
8.40 (dd, J' = 5.0 Hz, J'' = 1.3 Hz, 1H), 8.25 (d, J = 5.0 Hz, 1H),
7.22 (s, 2H), 7.18 (s, 1H), 7.14 (t, J = 9.0 Hz, 1H), 6.98 (d, J =
5.0 Hz, 1H), 6.61 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.54-6.51
(m, 1H), 3.84-3.82 (m, 1H), 1.85-1.82 (m, 2H), 1.75-1.72 (m, 2H),
1.62-1.59 (m, 1H), 1.48-1.30 (m, 4H), 1.21-1.15 (m, 1H) [M + H]
480.2 J 258 ##STR00575## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(3,5- dichlorophenyl) furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.28 (d, J = 1.6 Hz, 2H), 8.18 (d, J = 5.0
Hz, 1H), 7.71 (s, 1H), 7.19- 7.11 (m, 4H), 6.92 (d, J = 5.0 Hz,
1H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.5 Hz, 1H), 6.52-6.50 (m, 1H) [M
+ H] 422 A 259 ##STR00576## 7-(2-(tert- Butyl)pyridin-
4-yl)-N.sup.3-(3- chloro-4- fluorophenyl) furo[2,3-c] pyridine-2,3-
diamine DMSO-d.sub.6: .delta. 8.67 (d, J = 4.9 Hz, 1H), 8.24 (s,
1H), 8.22 (d, J = 4.9 Hz, 1H), 8.03 (d, J = 4.3 Hz, 1H), 7.18-7.11
(m, 4H), 6.94 (d, J = 4.9 Hz, 1H), 6.60-6.59 (m, 1H), 6.52-6.50 (m,
1H), 1.40 (s, 9H) [M + H] 411 A 260 ##STR00577## 2-Amino-3-((3-
chloro-4- fluorophenyl) amino)furo[2,3- c]pyridine 6- oxide
DMSO-d.sub.6: .delta. 8.43 (s, 1H), 7.83 (d, J = 6.4 Hz, 1H), 7.14
(t, J = 9.0 Hz, 2H), 6.99 (s, 2H), 6.77 (d, J = 6.5 Hz, 1H), 6.58
(dd, J' = 6.1 Hz, J'' = 2.5 Hz, 1H), 6.50-6.47 (m, 1H) [M + H]
294.1 A 261 ##STR00578## 7-(Benzo[d] [1,3]dioxol- 5-yl)-N.sup.3-
(3-chloro-4- fluorophenyl) furo[2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.11 (d, J = 5.2 Hz, 1H), 7.85 (d, J = 8.0
Hz, 1H), 7.80 (s, 1H), 7.15- 7.13 (m, 2H), 7.07 (d, J = 8.8 Hz,
1H), 7.01 (s, 2H), 6.81 (d, J = 5.2 Hz, 1H), 6.58 (dd, J' = 6.0 Hz,
J'' = 2.4 Hz, 1H), 6.52- 6.48 (m, 1H), 6.11 (s, 2H) [M + H] 398.1 A
262 ##STR00579## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(2-
ethylpyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine CD.sub.3CN:
.delta. 8.69 (d, J = 5.4 Hz, 1H), 8.20 (d, J = 5.5 Hz, 1H), 8.15
(s, 1H), 8.09 (d, J = 5.3 Hz, 1H), 7.11 (d, J = 5.4 Hz, 1H), 7.05
(t, J = 9.0 Hz, 1H), 6.66 (dd, J' = 6.2 Hz, 1H), 6.60 (dt, J' = 8.8
Hz, J'' = 6.5 Hz, J''' = 3.3 Hz, 1H), 6.04 (s, 2H), 5.80 (s, 1H),
2.98 (q, J = 7.6 Hz, 2H), 1.38 [M + H] 382.8 J (t, J = 7.6 Hz, 3H)
263 ##STR00580## N.sup.3-(5-Chloro-2- fluorophenyl)- 7-(2-
methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.61 (d, J = 5.2 Hz, 1H), 8.22 (d, J = 4.8 Hz, 1H), 8.10
(s, 1H), 8.04 (d, J = 4.8 Hz, 1H), 7.22 (d, J = 8.4 Hz, 2H), 7.15
(t, J = 10.2 Hz, 2H), 6.97 (d, J = 5.2 Hz, 1H), 6.64-6.62 (M, 1H),
6.29 (d, J = 6.0 Hz, 1H), 2.59 (s, 3H) [M + H] 369.2 A 264
##STR00581## N.sup.3-(3-Chloro-2- fluorophenyl)- 7-(2-
methylpyridin- 4-yl)furo[2,3- c]pyridine-2,3- diamine DMSO-d.sub.6:
.delta. 8.61 (d, J = 4.8 Hz, 1H), 8.20 (d, J = 5.2 Hz, 1H), 8.09
(s, 1H), 8.03 (d, J = 5.2 Hz, 1H), 7.18 (s, 3H), 6.96 (d, J = 5.2
Hz, 1H), 6.88 (t, J = 7.6 Hz, 1H), 6.74 (t, J = 6.8 Hz, 1H), 6.32
(t, J = 7.2 Hz, 1H), 2.67 (s, 3H) [M + H] 369.1 A 265 ##STR00582##
7-(2- Methylpyridin- 4-yl)-N.sup.3- phenylbenzo[b] thiophene-2,3-
diamine DMSO-d.sub.6: .delta. 8.52 (d, J = 5.0 Hz, 1H), 7.51 (s,
1H), 7.45-7.43 (m, 1H), 7.22 (t, J = 7.6 Hz, 1H), 7.08- 7.00 (m,
5H), 6.54 (t, J = 7.3 Hz, 1H), 6.49 (d, J = 7.8 Hz, 2H), 5.85 (s,
2H), 2.51 (s, 3H) [M + H] 332.12 I, E 266 ##STR00583## N.sup.3-(4-
Fluorophenyl)- 7-(2- methylpyridin- 4-yl)benzo[b] thiophene-2,3-
diamine DMSO-d.sub.6: .delta. 8.53 (d, J = 5.2 Hz, 1H), 7.51 (s,
1H), 7.44 (d, J = 4.6 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.08-7.06
(m, 2H), 6.99 (d, J = 7.8 Hz, 1H), 6.87 (t, J = 8.8 Hz, 2H), 6.46
(dd, J' = 8.7 Hz, J'' = 4.6 Hz, 2H), 5.90 (s, 2H), 2.51 (s, 3H) [M
+ H] 350.11 I, E 267 ##STR00584## N.sup.3-(3-Chloro-4-
fluorophenyl)- 5-methyl-7-(2- methylpyridin- 4-yl)furo[2,3-
c]pyridine-2,3- diamine DMSO-d.sub.6: .delta. 8.58 (d, J = 5.0 Hz,
1H), 8.06 (s, 1H), 8.01 (d, J = 4.6 Hz, 1H), 7.15- 7.12 (m, 2H),
7.07 (s, 2H), 6.78 (s, 1H), 6.59 (dd, J' = 6.2 Hz, J'' = 2.4 Hz,
1H), 6.50-6.48 (m, 1H), 2.58 (s, 3H), 2.45 (s, 3H) [M + H] 383.1 A
268 ##STR00585## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-(2-
(trifluoro- methyl) pyridin-4- yl)furo[2,3- c]pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.95 (d, J = 5.1 Hz, 1H), 8.64 (s, 1H), 8.55
(d, J = 4.8 Hz, 1H), 8.26 (d, J = 5.0 Hz, 1H), 7.28 (s, 2H), 7.19
(s, 1H), 7.14 (t, J = 9.1 Hz, 2H), 7.01 (d, J = 5.0 Hz, 1H), 6.61
(dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H) [M + H] 423.2 J 269
##STR00586## Ethyl (3-chloro- 4-fluorophenyl) (2-((ethoxy-
carbonyl) amino)-7-(2- methylpyridin- 4-yl)furo[2,3- c]pyridin-3-
yl)carbamate DMSO-d.sub.6: .delta. 8.67 (d, J = 5.2 Hz, 1H), 8.57
(d, J = 4.8 Hz, 1H), 8.35 (s, 1H), 8.07 (s, 1H), 8.02 (d, J = 5.2
Hz, 1H), 7.53 (d, J = 4.8 Hz, 1H), 7.26 (t, J = 9.0 Hz, 1H), 6.83
(dd, J' = 6.2 Hz, J'' = 2.8 Hz, 1H), 6.76- 6.73 (m, 1H), 4.17 (q, J
= 6.9 Hz, 4H), 2.60 (s, 3H), 1.10 (t, J = 7.0 Hz, 6H) [M + H] 513.2
H 271 ##STR00587## (4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo[2,3- c]pyridin-7- yl)pyridin-2- yl)(piperidin-1-
yl)methanone DMSO-d.sub.6: .delta. 8.87 (d, J = 5.2 Hz, 1H),
8.06-8.03 (m, 2H), 7.94 (d, J = 4.4 Hz, 1H), 7.21 (d, J = 6.4 Hz,
1H), 7.08 (t, J = 9.2 Hz, 1H), 6.91 (s, 2H), 6.69 (dd, J' = 6.0 Hz,
J'' = 2.8 Hz, 1H), 6.63 (dt, J' = 8.6 Hz, J'' = 7.0 Hz, J''' = 3.4
Hz, 1H), 5.90 (s, 1H), 3.71 (t, J = 5.2 Hz, 2H), 3.40 (t, J = 5.2
Hz, 2H), 1.71-1.66 (m, 4H), 1.57-1.55 (m, 2H) [M + H] 466.1 J 272
##STR00588## (4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo[2,3- c]pyridin-7- yl)pyridin-2- yl)(morpholino)
methanone DMSO-d.sub.6: .delta. 8.75 (d, J = 5.2 Hz, 1H), 8.40 (s,
1H), 8.33 (d, J = 5.0 Hz, 1H), 8.23 (d, J = 5.0 Hz, 1H), 7.22 (s,
2H), 7.18 (s, 1H), 7.14 (t, J = 9.1 Hz, 1H), 6.97 (d, J = 5.0 Hz,
1H), 6.60 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H), 6.51 (dt, J' = 8.8
Hz, J'' = 6.6 Hz, J''' = 3.2 Hz, 1H), 3.70 (s, 4H), 3.59 (d, J =
4.1 Hz, 2H), 3.50 (d, J = 4.1 Hz, 2H) [M + H] 468.3 J 273
##STR00589## 4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo[2,3- c]pyridin-7-yl)- N-(pyridin-2- yl)picolinamide
DMSO-d.sub.6: .delta. 10.51 (s, 1H), 9.05 (s, 1H), 8.93 (d, J = 5.1
Hz, 1H), 8.54 (d, J = 5.1 Hz, 1H), 8.43 (d, J = 4.2 Hz, 1H), 8.33
(d, J = 8.3 Hz, 1H), 8.28 (d, J = 5.0 Hz, 1H), 7.94 (t, J = 7.9 Hz,
1H), 7.31 (bs, 2H), 7.25-7.21 (m, 2H), 7.15 (t, J = 9.1 Hz, 1H),
7.01 (d, J = 5.1 Hz, 1H), 6.62 (dd, J' = 6.2 Hz, J'' = 2.6 Hz, 1H),
6.55-6.52 (m, 1H) [M + H] 475.3 J 274 ##STR00590## 4-(2-Amino-3-
((3-chloro-4- fluorophenyl) amino)furo[2,3- c]pyridin-7-yl)- N-(2-
morpholino- ethyl) picolinamide CD.sub.3CN: .delta. 9.01 (s, 1H),
8.74 (d, J = 5.0 Hz, 1H), 8.46-8.44 (m, 1H), 8.41 (bs, 1H), 8.19
(d, J = 4.9 Hz, 1H), 7.05-7.01 (m, 2H), 6.66 (dd, J' = 6.3 Hz, J''
= 2.7 Hz, 1H), 6.59 (dt, J' = 8.8 Hz, J'' = 6.7 Hz, J''' = 3.1 Hz,
1H), 5.74 (s, 1H), 5.61 (s, 2H), 3.67 (t, J = 4.4 Hz, 4H),
3.59-3.54 (m, 2H), 2.64-2.63 (m, 2H), 2.53 (s, 4H) [M + H] 511.2 J
275 ##STR00591## 4-(2-Amino-3- ((3-chloro-4- fluorophenyl)
amino)furo[2,3- c]pyridin-7-yl)- N-pentyl- picolinamide CD.sub.3CN:
.delta. 8.90 (s, 1H), 8.77 (d, J = 5.2 Hz, 1H), 8.33-8.32 (m, 1H),
8.24-8.20 (m, 2H), 7.10-7.02 (m, 2H), 6.67 (dd, J' = 6.0 Hz, J'' =
2.6 Hz, 1H), 6.60 (dt, J' = 8.8 Hz, J'' = 6.4 Hz, J''' = 3.2 Hz,
1H), 6.01 (s, 2H), 5.78 (s, 1H), 3.42 (q, J = 6.8 Hz, 2H),
1.65-1.61 (m, 2H), 1.38-1.36 (m, 4H), 0.91 (t, J = 6.8 Hz, 3H) [M +
H] 468.2 J 276 ##STR00592## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(2-chloro-5- fluorophenyl) furo[2,3- c]pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.22 (d, J = 4.8 Hz, 1H), 7.60-7.57 (m, 1H),
7.36 (d, J = 6.5 Hz, 1H), 7.27-7.23 (m, 1H), 7.06-7.02 (m, 2H),
6.65-6.64 (m, 1H), 6.58-6.56 (m, 1H), 5.73 (s, 1H), 5.40 (s, 2H) [M
+ H] 406.1 A 277 ##STR00593## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(3,5- dimethyl- phenyl)furo [2,3-c] pyridine-2,3- diamine
DMSO-d.sub.6: .delta. 8.12 9 (d, J = 5.1 Hz, 1H), 7.23-7.21 (m,
2H), 7.17-7.12 (m, 3H), 6.93 (bs, 2H), 6.87 (d, J = 5.0 Hz, 1H),
6.61-6.59 (m, 1H), 6.51-6.48 (m, 1H), 2.32 (s, 3H), 2.16 (s, 3H) [M
+ H] 381.8 A 278 ##STR00594## N.sup.3-(3-Chloro-4- fluorophenyl)-
7-(3,5- difluorophenyl) furo[2,3-c] pyridine-2,3- diamine
CD.sub.3CN: .delta. 8.22 (d, J = 4.7 Hz, 1H), 8.01 (d, J = 8.8 Hz,
2H), 7.05-6.99 (m, 3H), 6.67-6.64 (m, 1H), 6.59-6.57 (m, 1H), 5.73
(s, 1H), 5.59 (s, 2H) [M + H] 390.2 A 279 ##STR00595## Ethyl (3,4-
difluorophenyl) (2-((ethoxy- carbonyl) amino)-7-(2- methylpyridin-
4-yl)furo[2,3- c]pyridin-3- yl)carbamate DMSO-d.sub.6: .delta. 8.72
(bs, 1H), 8.57 (d, J = 4.8 Hz, 1H), 8.33 (s, 1H), 8.09 (s, 1H),
8.03 (s, 1H), 7.51 (d, J = 4.8 Hz, 1H), 7.30-7.23 (m, 1H),
6.70-6.66 (m, 1H), 6.56-6.54 (m, 1H), 4.17 (q, J = 7.1 Hz, 4H),
2.60 (s, 3H), 1.09 (t, J = 7.0 Hz, 6H) [M + H] 497.3 H 280
##STR00596## N.sup.3-(3-Chloro-4- fluorophenyl)- 7-styrylfuro
[2,3-c]pyridine- 2,3-diamine DMSO-d.sub.6: .delta. 8.08 (d, J = 5.2
Hz, 1H), 7.81 (d, J = 16.0 Hz, 1H), 7.67 (d, J = 7.6 Hz, 2H),
7.45-7.39 (m, 3H), 7.34 (t, J = 7.2 Hz, 1H), 7.13 (t, J = 8.8 Hz,
2H), 6.98 (s, 2H), 6.78 (d, J = 5.2 Hz, 1H), 6.58 (dd, J' = 6.0 Hz,
J'' = 2.6 Hz, 1H), 6.50 (dt, J' = 8.8 Hz, J'' = 6.8 Hz, J'' = 3.6
Hz, 1H), [M + H] 380.2 A 281 ##STR00597## N.sup.3-(3-Chloro-4-
fluorophenyl)- 7-(isoquinolin- 4-yl)benzo[b] thiophene-2,3- diamine
DMSO-d.sub.6: .delta. 9.38 (s, 1H), 8.53 (s, 1H), 8.22 (d, J = 5.3
Hz, 1H), 7.71 (s, 2H), 7.58 (s, 1H), 7.32- 7.29 (m, 2H), 7.13- 7.07
(m, 2H), 7.03 (d, J = 6.9 Hz, 1H), 6.57-6.56 (m, 1H), 6.48-6.46 (m,
1H), 5.89 (s, 2H), [M + H] 420.07 I
[0769] TABLE 2. is a non-exhaustive list of compounds of the
invention that can be made using according the procedures described
herein.
TABLE-US-00005 TABLE 2 No. Structure IUPAC Name M + 1 301
##STR00598## N.sup.3-(3-Chloro-4- fluorophenyl)-7-(pyridin-2-
yl)furo[2,3-c]pyridine-2,3- diamine 355.76 302 ##STR00599##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)-7-
(pyridine-2-yl)furo[2,3- c]pyridine-5-carbonitrile 380.06 303
##STR00600## 2-Amino-3-((3-chloro-4- fluorophenyl)amino)-7-
phenylfuro[2,3-c]pyridine-5- carbonitrile 379.07 304 ##STR00601##
N.sup.3-(3-Methoxy-5- methylphenyl)furo[2,3- c]pyridine-2,3-diamine
270.29 305 ##STR00602## N.sup.7-(3-Chloro-4- fluorophenyl)furo[3,2-
d]pyrimidine-6,7-diamine 279.67 306 ##STR00603##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridine-7-carboxylic acid 322.69 307 ##STR00604##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridine-7-carboxamide 321.70 308 ##STR00605##
N.sup.3-(3-Chloro-4- fluorophenyl)-N5,N5-
dimethyl-7-phenylfuro[2,3- c]pyridine-2,3,5-triamine 397.84 309
##STR00606## N.sup.3-(3- Cyclobutylphenyl)furo[2,3-
c]pyridine-2,3-diamine 280.33 310 ##STR00607## N.sup.3-(3-
Cyclopentylphenyl)furo[2,3- c]pyridine-2,3-diamine 294.36 311
##STR00608## N.sup.3-(3-Chloro-4- fluorophenyl)-7-
(trifluoromethyl)furo[2,3- c]pyridine-2,3-diamine 346.67 312
##STR00609## N.sup.3-(3-Chloro-4- fluorophenyl)-1H-pyrrolo[2,3-
c]pyridine-2,3-diamine 276.68 313 ##STR00610## N.sup.3-(3-Chloro-4-
fluorophenyl)-7- phenoxyfuro[2,3-c]pyridine- 2,3-diamine 370.07 314
##STR00611## N.sup.3-(3-Chloro-4- fluorophenyl)-7-(pyridine-2-
yloxy)furo[2,3-c]pyridine-2,3- diamine 371.06 315 ##STR00612##
N.sup.3-(3-Chloro-4- fluorophenyl)-N.sup.7-
phenylfuro[2,3-c]pyridine- 2,3,7-triamine 369.08 316 ##STR00613##
N.sup.3-(3-Chloro-4- fluorophenyl)-N.sup.7-(pyridine-3-
yl)furo[2,3-c]pyridine-2,3,7- triamine 370.08 317 ##STR00614##
N.sup.3-(3-Chloro-4- fluorophenyl)-7- (phenylethynyl)furo[2,3-
c]pyridine-2,3-diamine 378.07 318 ##STR00615## N.sup.3-(3-Chloro-4-
fluorophenyl)-N.sup.7-(pyridine-3- yl)furo[2,3-c]pyridine-2,3,7-
triamine 370.08 319 ##STR00616## N.sup.3-(3-Chloro-4-
fluorophenyl)-5-(2- (dimethylamino)ethoxy)-7-
(pyridine-2-yl)furo[2,3- c]pyridine-2,3-diamine 442.14 320
##STR00617## N.sup.3-(3-Chloro-4- fluorophenyl)-5-(3-
dimethylamino)ethoxy)-7- phenylfuro[2,3-c]pyridine- 2,3-diamine
441.14 321 ##STR00618## N.sup.3-(3-Chloro-4-
fluorophenyl)-N.sup.5,N.sup.5- dimethyl-7-phenylfuro[2,3-
c]pyridine-2,3,5-triamine 397.12 322 ##STR00619##
N.sup.3-(3-Chloro-4- fluorophenyl)-7-(1-methyl-
1H-pyrazol-5-yl)furo[2,3- c]pyridine-2,3-diamine 358.08 323
##STR00620## N.sup.3-(3-Chloro-4- fluorophenyl)-7-(piperidin-1-
yl)furo[2,3-c]pyridine-2,3- diamine 361.12 324 ##STR00621##
N.sup.3-(3-Chloro-4- fluorophenyl)-7-(4- methylpiperazin-1-
yl)furo[2,3-c]pyridine-2,3- diamine 376.13 325 ##STR00622##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)-N-
phenylfuro[2,3-c]pyridine-7- carboxamide 397.08 326 ##STR00623##
N.sup.3-(3-Bromo-4- methylphenyl)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 409.06 327
##STR00624## N.sup.3-([1,1'-Biphenyl]-3-yl)-7- (2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 393.17 328 ##STR00625##
N.sup.3-(3-Bromo-4- fluorophenyl)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 413.04 329
##STR00626## N.sup.3-(3-(1,1-Difluoroethyl)-4- fluorophenyl)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 399.14 330
##STR00627## N.sup.3-(3-Chloro-2,4- difluorophenyl)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 387.08 331
##STR00628## 7-(2-Methylpyridin-4-yl)-N.sup.3- (4-
(trifluoromethyl)phenyl)furo [2,3-c]pyridine-2,3-diamine 385.12 332
##STR00629## 7-(2-Methylpyridin-4-yl)-N.sup.3- (4-
(trifluoromethoxy)phenyl)furo [2,3-c]pyridine-2,3-diamine 401.12
333 ##STR00630## 7-(2-Methylpyridin-4-yl)-N.sup.3-
(pyridin-3-yl)furo[2,3- c]pyridine-2,3-diamine 318.13 334
##STR00631## N.sup.3-(4-Fluoro-3- (trifluoromethoxy)phenyl)-7-
(2-methylpyridin-4- yl)furo[2,3-c]pyridine-2,3- diamine 419.11 335
##STR00632## N.sup.3-(3- (Difluoromethyl)phenyl)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 367.13 336
##STR00633## 3-((2-Amino-7-(2- methylpyridin-4-yl)furo[2,3-
c]pyridin-3- yl)amino)benzonitrile 342.13 337 ##STR00634##
N.sup.3-(4-Chlorophenyl)-7-(2- methylpyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 351.10 338 ##STR00635##
N.sup.3-Hexyl-7-(2-methylpyridin- 4-yl)furo[2,3-c]pyridine-2,3-
diamine 325.20 339 ##STR00636## N.sup.3-Benzyl-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-2,3-diamine 331.15 340
##STR00637## N.sup.3-Cyclohexyl-7-(2- methylpyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 323.18 341 ##STR00638##
7-(2-Methylpyridin-4-yl)-N.sup.3- (tetrahydro-2H-pyran-4-
yl)furo[2,3-c]pyridine-2,3- diamine 325.16 342 ##STR00639##
7-(2-Methylpyridin-4-yl)-N.sup.3- ((tetrahydro-2H-pyran-4-
yl)methyl)furo[2,3-c]pyridine- 2,3-diamine 339.18 343 ##STR00640##
(E)-N.sup.3-(4-Fluorophenyl)-7- styrylfuro[2,3-c]pyridine-2,3-
diamine 346.13 344 ##STR00641## (E)-N.sup.3-(3,4-Difluorophenyl)-
7-styrylfuro[2,3-c]pyridine- 2,3-diamine 364.12 345 ##STR00642##
(E)-N.sup.3-(4-Fluoro-3- (trifluoromethyl)phenyl)-7-
styrylfuro[2,3-c]pyridine-2,3- diamine 414.12 346 ##STR00643##
2-Amino-3-((3,4- difluorophenyl)amino)-N-
methylfuro[2,3-c]pyridine-7- carboxamide 319.10 347 ##STR00644##
7-(Aminomethyl)-N.sup.3-(3- chlorophenyl)furo[2,3-
c]pyridine-2,3-diamine 289.08 348 ##STR00645##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3- c]pyridin-7-yl
acetate 336.05 349 ##STR00646## N-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)acetamide 335.07 350
##STR00647## N.sup.3-(2-Chlorophenyl)-7- (piperidin-4-
ylmethyl)furo[2,3-c]pyridine- 2,3-diamine 357.14 351 ##STR00648##
N.sup.3-(3,4-Difluorophenyl)-7- (pyridin-4-ylmethyl)furo[2,3-
c]pyridine-2,3-diamine 353.12 352 ##STR00649## (2-Amino-3-((3,4-
difluorophenyl)amino)furo[2, 3-c]pyridin-7-yl)methanol 292.08 353
##STR00650## N.sup.3-(3-Chlorophenyl)-7- (trifluoromethyl)furo[2,3-
c]pyridine-2,3-diamine 328.04 354 ##STR00651## N.sup.3-(3-Chloro-4-
fluorophenyl)-7- (cyclohexyloxy)furo[2,3- c]pyridine-2,3-diamine
376.12 355 ##STR00652## N.sup.3-(3,4-Difluorophenyl)-7-
((tetrahydro-2H-pyran-4- yl)oxy)furo[2,3-c]pyridine- 2,3-diamine
362.13 356 ##STR00653## 2-Amino-3-((3-chloro-4-
fluorophenyl)amino)-N- methylfuro[2,3-c]pyridine-7- sulfonamide
371.03 357 ##STR00654## 2-Amino-3-((3,4- difluorophenyl)amino)-N-
phenylfuro[2,3-c]pyridine-7- sulfonamide 417.08 358 ##STR00655##
N.sup.3-(4-Fluorophenyl)-5- (trifluoromethyl)furo[2,3-
c]pyridine-2,3-diamine 312.07 359 ##STR00656##
N.sup.3-(2-Chlorophenyl)-5- (trifluoromethoxy)furo[2,3-
c]pyridine-2,3-diamine 344.04 360 ##STR00657##
2-Amino-3-((3-chloro-4- fluorophenyl)amino)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridine-5-carbonitrile 394.08 361
##STR00658## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)-N,N-
dimethylpicolinamide 426.11 362 ##STR00659##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-N- ethylpicolinamide 426.11 363 ##STR00660##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-N- isopropylpicolinamide 440.12 364 ##STR00661##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-N-(2- methoxyethyl)picolinamide 456.12 365
##STR00662## N-(3-(1H-Imidazol-1- yl)propyl)-4-(2-amino-3-((3-
chloro-4- fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)picolinamide
506.15 366 ##STR00663## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)-N-(pyridin-3-
yl)picolinamide 475.10 367 ##STR00664## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)-N-(pyridin-4-
yl)picolinamide 475.10 368 ##STR00665## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)-N-(4-
methylpiperazin-1- yl)picolinamide 496.16 369 ##STR00666##
N.sup.3-(3-Chloro-4- fluorophenyl)thieno[3,2-
b]pyridine-2,3-diamine 294.02 370 ##STR00667## 3-((3-Chloro-4-
fluorophenyl)thio)furo[2,3- c]pyridin-2-amine 295.0 371
##STR00668## 3-((3-Chloro-4- fluorophenyl)thio)benzofuran- 2-amine
294.0 372 ##STR00669## 3-(3-Chloro-4- fluorophenoxy)furo[2,3-
c]pyridin-2-amine 279.0 373 ##STR00670## 3-(3-Chloro-4-
fluorophenoxy)benzofuran-2- amine 278.0 374 ##STR00671##
N3-(3-Chloro-4- fluorophenyl)benzofuran-2,3- diamine 277.05 375
##STR00672## N3-(3-Chloro-4- fluorophenyl)-7-(2- methylpyridin-4-
yl)benzofuran-2,3-diamine 368.09 376 ##STR00673## N3-(3-Chloro-4-
fluorophenyl)-7-(2,6- difluoropyridin-4- yl)benzofuran-2,3-diamine
390.06 377 ##STR00674## N3-(3-Chloro-4- fluorophenyl)-7-(2,6-
difluoropyridin-4- yl)benzo[b]thiophene-2,3- diamine 406.03 378
##STR00675## N3-(3-Chloro-4- fluorophenyl)-7-(2- fluoropyridin-4-
yl)benzofuran-2,3-diamine 372.07 379 ##STR00676## N3-(3-Chloro-4-
fluorophenyl)-7-(2- fluoropyridin-4- yl)benzo[b]thiophene-2,3-
diamine 388.04 380 ##STR00677## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)benzofuran- 7-yl)-N- methylpicolinamide 411.10
381 ##STR00678## 2-Amino-3-((3-chloro-4- fluorophenyl)amino)-7-(2-
methylpyridin-4-yl)furo[2,3- c]pyridin-5-ol 385.08 382 ##STR00679##
N.sup.3-(3-Chloro-4- fluorophenyl)-7-(2-
fluoropyridin-4- yl)benzofuran-2,3-diamine 373.06 383 ##STR00680##
N3-(3-chlorophenyl)-7-(2- fluoropyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 255.07 384 ##STR00681##
N3-(3-Fluorophenyl)-7-(2- fluoropyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 339.10 385 ##STR00682##
N3-(3,4-Difluorophenyl)-7- (2-fluoropyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 357.09 386 ##STR00683##
N3-(4-Fluorophenyl)-7-(2- fluoropyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 339.10 387 ##STR00684## N3-(3-Chloro-4-
fluorophenyl)-1H-indole-2,3- diamine 276.07 388 ##STR00685##
N3-(3-Chloro-4- fluorophenyl)-7-(2- methylpyridin-4-yl)-1H-
pyrrolo[2,3-c]pyridine-2,3- diamine 368.10 389 ##STR00686##
N3-(3-Chloro-4- fluorophenyl)-7-(2- methylpyridin-4-yl)-1H-
indole-2,3-diamine 367.11 390 ##STR00687##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)-1H-
pyrrolo[2,3-c]pyridin-7-yl)-N- methylpicolinamide 411.11 391
##STR00688## 4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)-1H-
indol-7-yl)-N- methylpicolinamide 410.11 392 ##STR00689##
N3-(3-Chloro-4- fluorophenyl)-7-(4- methylpyrimidin-2-
yl)furo[2,3-c]pyridine-2,3- diamine 370.08 393 ##STR00690##
N3-(3-Chloro-4- fluorophenyl)-7-(imidazo[1,2-
a]pyridin-5-yl)furo[2,3- c]pyridine-2,3-diamine 394.08 394
##STR00691## N3-(3-Chloro-4- fluorophenyl)-7-(imidazo[1,2-
a]pyridin-2-yl)furo[2,3- c]pyridine-2,3-diamine 394.08 395
##STR00692## 4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3
- c]pyridin-7-yl)-N-methyl-1H- imidazole-1-carboxamide 401.09 396
##STR00693## N3-(1-Methyl-1H-pyrrol-3- yl)-7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 320.15 397 ##STR00694##
7-(2-Methylpyridin-4-yl)-N3- (1H-pyrrol-3-yl)furo[2,3-
c]pyridine-2,3-diamine 306.13 398 ##STR00695##
N3-(1-Methyl-1H-pyrrol-2- yl)-7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 320.15 399 ##STR00696##
N3-(1-Methyl-1H-indol-2-yl)- 7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 370.16 400 ##STR00697##
N3-(6-Chloropyrimidin-4-yl)- 7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 353.09 401 ##STR00698##
N3-(6-Fluoropyrimidin-4-yl)- 7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 337.12 402 ##STR00699##
N3-(2-Fluoropyrimidin-5-yl)- 7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 337.12 403 ##STR00700##
N3-(4,5-difluoropyrimidin-2- yl)-7-(2-methylpyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 355.11 404 ##STR00701##
N.sup.3-(3-Chlorophenyl)-7-(2,6- difluoropyridin-4-yl)furo[2,3-
c]pyridine-2,3-diamine 373.06 405 ##STR00702##
N3-(3,4-Difluorophenyl)-7- (2,6-difluoropyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 375.08 406 ##STR00703##
7-(2,6-Difluoropyridin-4-yl)- N3-(4-fluorophenyl)furo[2,3-
c]pyridine-2,3-diamine 357.09 407 ##STR00704##
N3-(5-chloro-2-fluorophenyl)- 7-(2,6-difluoropyridin-4-
yl)furo[2,3-c]pyridine-2,3- diamine 391.05 408 ##STR00705##
N3-(3,4-Difluorophenyl)-7- (4- (methoxymethyl)phenyl)furo
[2,3-c]pyridine-2,3-diamine 382.13 409 ##STR00706##
7-(2-Chloro-6-fluorophenyl)- N3-(3,4- difluorophenyl)furo[2,3-
c]pyridine-2,3-diamine 403.06 410 ##STR00707##
N-(4-(2-Amino-3-((3,4- difluorophenyl)amino)furo[2, 3-c]pyridin-7-
yl)phenyl)pyrrolidine-1- carboxamide 450.17 411 ##STR00708##
N3-(3,4-Difluorophenyl)-7- (4-fluoro-2- methylphenyl)furo[2,3-
c]pyridine-2,3-diamine 370.11 412 ##STR00709##
N3-(3,4-Difluorophenyl)-7- (4-propoxyphenyl)furo[2,3-
c]pyridine-2,3-diamine 396.15 413 ##STR00710##
N3-(3,4-Difluorophenyl)-7- (4- morpholinophenyl)furo[2,3-
c]pyridine-2,3-diamine 423.16 414 ##STR00711##
5-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-2-fluorophenol 388.06 415 ##STR00712## Methyl
5-(2-amino-3-((3- chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-2- chlorobenzoate 446.04 416 ##STR00713##
3-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-N- methylbenzamide 411.10 417 ##STR00714##
4-(4-(2-Amino-3-((3-chloro- 4- fluorophenyl)amino)furo[2,3-
c]pyridin-7- yl)picolinamido)butanoic acid 484.11 418 ##STR00715##
7-(2,6-Difluoropyridin-4-yl)- N.sup.3-(4-
(trifluoromethyl)phenyl)furo [2,3-c]pyridine-2,3-diamine 407.09 419
##STR00716## 7-(2-Fluoropyridin-4-yl)-N.sup.3- (4-
(trifluoromethyl)phenyl)furo [2,3-c]pyridine-2,3-diamine 389.10 420
##STR00717## 5-Fluoro-7-(2-methylpyridin- 4-yl)-N3-(4-
(trifluoromethyl)phenyl)furo [2,3-c]pyridine-2,3-diamine 403.11 421
##STR00718## 4-(2-Amino-3-((4- (trifluoromethyl)phenyl)amino)
furo[2,3-c]pyridin-7-yl)-N- methylpicolinamide 428.13 422
##STR00719## 4-(2-Amino-5-fluoro-3-((4-
(trifluoromethyl)phenyl)amino) furo[2,3-c]pyridin-7-yl)-N-
methylpicolinamide 446.12 423 ##STR00720##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)-5-
fluorofuro[2,3-c]pyridin-7- yl)-N-methylpicolinamide 430.08 424
##STR00721## 5-Fluoro-N.sup.3-(4- (trifluoromethyl)phenyl)furo
[2,3-c]pyridine-2,3-diamine 312.07 425 ##STR00722##
N.sup.3-(3-Chloro-4- fluorophenyl)-6- fluorobenzo[b]thiophene-2,3-
diamine 311.02 426 ##STR00723## N.sup.3-(3-Chloro-4-
fluorophenyl)-1-methyl-1H- indole-2,3-diamine 290.08 427
##STR00724## 7-(2,6-difluoropyridin-4-yl)-
N3-(3-fluorophenyl)furo[2,3- c]pyridine-2,3-diamine 357.1 428
##STR00725## N.sup.3-(2,5-Ddifluorophenyl)-7-
(2,6-difluoropyridin-4- yl)furo[2,3-c]pyridine-2,3- diamine 375.09
429 ##STR00726## 4-(2-Amino-3-((3-chloro-4-
fluorophenyl)amino)furo[2,3- c]pyridin-7-yl)-N-(2-
hydroxyethyl)picolinamide 442.11 430 ##STR00727##
4-(2-Amino-3-((3-chloro-4- fluorophenyl)amino)furo[2,3-
c]pyridin-7-yl)-N,N- diethylpicolinamide 454.14 431 ##STR00728##
N.sup.3-(3-Chloro-4- fluorophenyl)-7-(2,3- dichlorophenyl)furo[2,3-
c]pyridine-2,3-diamine 422.00 432 ##STR00729## N.sup.3-(3-Chloro-4-
fluorophenyl)-7-(naphthalen- 2-yl)benzo[b]thiophene-2,3- diamine
419.08
[0770] One of skill in the art will recognize that the compounds of
the invention may be made as described herein and by methods known
in the art.
Example 30
[0771] A. In-vitro IDO1 Enzyme (Indoleamine 2,3-dioxygenase)
Assay
[0772] Human indoleamine 2,3-dioxygenasel (hIDO1) catalyzes the
oxidative cleavage of the pyrrole ring of the indole nucleus of
tryptophan to yield N-formylkynurenine which can be converted to
kynurenine (KYN) by deformylation. hIDO1 with an N-terminal His
tag, expressed and purified from E. coli cells was procured from
either Enzo LifeScienccs, NY, USA). Unless otherwise stated, all
materials were procured from Sigma Aldrich, Mo., USA.
[0773] The assay monitoring the conversion of L-tryptophan to KYN
was carried out as follows. hIDO1 (10 nM) was incubated with
tryptophan (30 .mu.M) in the presence of ascorbic acid (20 mM),
methylene blue (10 .mu.M) and catalase (100 .mu.g/mL) in potassium
phosphate buffer (50 mM; pH 6.5) at 37C for 30 min. The reaction
was terminated with 30% trichloroacetic acid (TCA) and further
incubated at 65C for 15 min to fully convert N-formylkynurenine to
KYN. The reaction mixture was then centrifuged to remove sediments
and the KYN in the supernatant was estimated by UV-visible
absorption spectroscopy at 360 nm using a Waters HPLC system fitted
with a C-18 column or by LC/MS/MS. See, Sono, 1980, J. Biol. Chem.,
255:1339-1345, which is herein incorporated by reference.
[0774] Percent inhibition at each concentration of test compounds
was determined by estimating the decrease in KYN with reference to
the reaction control with 1% DMSO vehicle. Data were analyzed using
nonlinear regression to generate IC50 values using Graph Pad
Prism.RTM. 5. The test compounds inhibit IDO1 activity and reduce
the levels of kynurenine pathway metabolite KYN as shown below in
Table 3.
B. HeLa Cell Based IDO1 (Indoleamine 2,3-dioxygenase) Assay
[0775] HeLa cells were obtained from ATCC and maintained in DMEM
supplemented with sodium bicarbonate (2.1 g/L), HEPES (4.1 g/L),
L-glutamine (2 mM), non-essential amino acid (84 mg/L) and fetal
bovine serum (10% FBS) and maintained at 95% humidity and 5%
CO.sub.2 in a 37.degree. C. incubator. Unless otherwise stated, all
materials were procured from Sigma. Upon incubation with
gamma-interferon (IFN.gamma. ), HeLa cells express IDO1, which
catalyzes the formation of N-formyl kynurenine from tryptophan
present in growth medium. The assay was performed as follows:
[0776] Cells were plated in medium (300 .mu.L) at a density of 0.1
million per well of a 48 well plate and hIDO1 was induced by
overnight treatment with IFN.gamma. (50 ng/mL) (Peprotech, USA).
The following day, cells were washed to remove IFN.gamma. and were
incubated with specific concentrations of test compounds (typically
10 .mu.M to 1 nM, final volume 300 .mu.L) in Hanks Balanced Salt
Solution (HBSS) containing 80 .mu.M L-tryptophan. Following
incubation, supernatant (150 .mu.L) was transferred to a 96 well
plate into which 30%TCA (304) was added and the contents further
incubated at 65.degree. C. for 15 min to fully convert
N-formylkynurenine to kynurenine. The reaction mixture was then
centrifuged to remove sediments and the KYN in the supernatant was
estimated by UV-visible absorption spectroscopy at 360 nm using a
Waters HPLC system fitted with a C-18 column. (Xiangdong Liu et al.
Blood 2010, Vol-117, 3520-30; Sono described above).
[0777] Percent inhibition at each concentration of test compounds
was determined by estimating the decrease in KYN with reference to
the reaction control with 1% DMSO vehicle. Data were analyzed using
nonlinear regression to generate IC50 values using Graph Pad
Prism.RTM. 5. The test compounds inhibit IDO1 and reduce kynurenine
pathway metabolite KYN (Table 3).
TABLE-US-00006 TABLE 3 A: IC50 < 200 nM; B: IC50 = 200 to 1000
nM; C: IC50 > 1000 nM. Enz = IDO1 Enzyme assay, Cell = HeLa cell
assay IC.sub.50 (IDO1) No. Enz Cell 1 A 2 A A 3 B A 4 C A 5 B A 6 C
7 C 8 B A 9 C B 10 C B 11 C 12 C 13 B A 14 B A 15 B B 16 C B 17 C
18 C 19 B B 20 A A 21 B A 22 C B 23 C 24 C 25 C B 26 A A 27 A A 28
C 29 C 30 C 31 C 32 C 33 A A 34 C 35 B A 36 C 37 C 38 C 39 C 40 C A
41 C 42 A A 43 B A 44 B A 45 C 46 B A 47 C B 48 A A 49 C 50 C 51 C
A 52 C 53 C 54 C 55 C A 56 C 57 C 58 A A 59 B A 60 A A 61 C 62 B A
63 C 64 A A 65 A A 66 A A 67 C A 68 B A 69 B A 70 B A 71 C A 72 B B
73 A A 74 A A 75 C 76 A A 77 A A 78 A A 79 A A 80 A A 81 A A 82 A A
83 A A 84 C B 85 A A 86 A A 87 B B 88 C 89 C 90 A A 91 A A 92 A A
93 B A 94 A A 95 A A 96 A A 97 A A 98 A A 99 C A 100 A A 101 A A
102 C C 103 A A 104 C 105 B C 106 C C 107 C 108 A A 109 B B 110 A A
111 A A 112 A A 113 A A 114 B A 115 A A 116 C B 117 A A 118 B A 119
A A 120 A A 121 C 122 C 123 A B 124 A A 125 C 126 B B 127 C 128 A
129 B 130 B B 131 B B 132 B B 133 B B 134 C 135 C 136 C 137 B A 138
C B 139 C C 140 C 141 C 142 C 143 C 144 C 145 C 146 C 147 A A 148 A
A 149 A A 151 A A 152 A A 153 A A 154 A A 155 C 156 C 157 C 158 A A
159 A A 160 C 161 C 162 C 163 C 164 A A 165 C 166 A A 167 A A 169 A
A 170 A A 171 A 172 A A 173 A A 174 A B 175 A A 176 A A 177 A A 179
A A 180 A A 181 A A 182 A A 183 A A 184 A A 185 B A 186 C C 187 B A
188 A A 189 A A 190 A A 191 A A 192 A A 193 C 194 C 195 A A 196 B A
197 A A 198 A A 199 A A 200 B A 201 A A 202 A A 203 A A 204 A A 205
A A 206 A A 207 A A 208 A A 209 C 210 A B 211 A A 212 A A 213 A A
214 B A 215 B A 216 A A 217 A A 218 B A 219 A A 220 A A 221 A A 222
A A 223 A A 224 A A 225 A A 226 A A 227 A A 228 A A 229 A A 230 A A
231 B B 232 C C 233 A A 234 A A 235 B A 236 A A 237 C B 238 A A 239
A A 240 A A 241 A A 242 C 243 A A 244 A A 245 A A 246 A A
247 A A 248 A A 249 A B 250 A A 251 A A 252 A A 253 A A 254 A A 256
A A 257 A A 258 A B 259 A B 260 B B 261 A A 262 A A 263 A A 264 B A
265 A 266 A 267 A A 268 A A 269 C C 271 A A 272 A A 273 A A 274 A A
275 A B 276 A A 277 A A 278 A A 279 C C 280 A A
C. CHO-K1 Cell Based TDO (Tryptophan 2,3-dioxygenase) Assay
[0778] Human TDO (hTDO)-transfected CHO-K1 cells were used to
identify test compounds that inhibit TDO activity and reduce
Kynurenine production. CHO-K1 cells (ATCC) were cultured in DMEM
supplemented with sodium bicarbonate (2.1 g/L), HEPES (4.1 g/L),
L-glutamine (2 mM), non-essential amino acid (84 mg/L) and fetal
bovine serum (10%) and maintained at 95% humidity and 5% CO.sub.2
in a 37.degree. C. incubator. The TDO gene (TDO2) (Accession number
NM_005651.1) was purchased from Origene (USA) and stably expressed
in CHO-Kl cells using the expression vector pcDNA4/Myc-HisB Hygro
(Life Technologies, USA). TDO-expressing clones were identified
based on the kynurenine production by the cells. Expanded clones
were used for assay as described below. Unless otherwise stated,
all materials were procured from Sigma.
[0779] hTDO transfected CHO-K1 cells (0.05 million/well) were
seeded in a 96-well plate and maintained overnight in DMEM
containing 10% FBS (200 .mu.L) at 95% humidity and 5% CO.sub.2 in a
37.degree. C. incubator. The following day, cells were washed and
incubated with test compounds (typically 31 .mu.M to 10 nM, final
volume 150 .mu.L) in Hanks Balanced Salt Solution (HBSS) containing
80 .mu.M L-tryptophan for 2 hours. Supernatant (150 .mu.L) from the
cells was treated with 30% TCA (30nL) to precipitate proteins.
Supernatants were further incubated at 65.degree. C. for 15 min to
convert N-formylkynurenine to kynurenine. KYN in the supernatant
was measured by LC/MS/MS using an API4000 mass spectrometer
(Applied Biosystems) coupled to a Shimadzu Prominence LC system
fitted with a C18 column.
[0780] Percent inhibition of the test compounds was determined by
measuring the decrease in KYN as compared to the reaction control
(DMSO vehicle). IC.sub.50 values were calculated using Graph Pad
Prism.RTM. 5 as described above. Compounds inhibited TDO activity
in cell culture system and reduced the levels of kynurenine
metabolite KYN as shown below in Table 4.
TABLE-US-00007 TABLE 4 A: IC50 < 500 nM; B: IC50 = 500 to 2000
nM; C: IC50 > 2000 nM hTDO cellular assay No. IC.sub.50 (TDO) 2
B 3 A 4 C 5 B 6 B 7 C 8 B 9 B 10 C 11 C 12 C 13 B 14 B 15 B 16 B 17
B 18 B 19 C 20 B 21 B 22 C 23 B 24 C 25 B 26 B 27 B 28 C 29 C 30 B
31 C 32 B 33 B 34 C 35 B 36 B 37 C 39 B 40 B 41 B 42 C 43 C 44 B 45
C 46 B 47 B 49 B 50 B 51 C 52 B 53 C 54 B 55 C 56 C 57 B 58 A 59 B
60 B 61 C 62 C 63 C 64 B 65 B 66 A 67 B 68 B 69 C 70 B 71 C 72 B 73
A 74 C 75 C 76 B 77 C 78 C 79 B 80 C 81 A 82 C 83 B 84 C 85 B 87 B
88 B 89 C 90 B 93 B 94 A 95 A 96 B 97 A 98 B 99 C 100 C 102 C 103 A
104 C 105 C 106 B 107 C 108 B 109 C 110 B 111 B 113 B 114 C 115 A
116 C 117 A 118 A 119 C 120 A 121 C 122 C 123 C 124 C 125 C 126 C
127 C 128 C 129 C 130 C 131 B 132 C 133 B 135 C 136 C 137 C 138 B
139 B 141 C 142 C 143 C 144 C 145 C 146 C 147 B 148 B 149 B 151 B
152 B 153 A 154 C 155 B 156 C 157 C 158 B 159 A 160 C 161 C 162 C
163 C 164 C 165 C 166 A 167 C 172 C 173 C 174 B 175 C 176 A 177 B
179 B 180 B 181 A 182 C 183 B 184 A 185 A 186 B 187 A 188 B 189 B
190 B 191 B 192 B 193 C 194 C 195 B 196 A 197 A 198 A 199 C 200 C
201 A 202 B 203 B 204 A 205 B 206 B 207 B 208 A 209 C 210 B 211 A
213 B 214 C 215 C 216 A 217 A 218 B 219 B 220 A 221 B 222 A 223 B
224 A 225 B 226 C 227 C 228 A 229 B 230 B 231 C 232 C 233 C 234 A
235 A 236 C 237 C 238 B 239 A 240 C 241 A 242 C 243 A 244 B 245 B
246 B 247 C 248 B 249 C 250 C 251 C 252 C 253 C 254 B 256 A 257 C
258 C 259 C 260 B 261 B 262 B
263 C 264 C 265 B 266 B 267 C 268 C 269 C 271 B 272 B 273 C 274 B
275 C 276 B 277 B 278 B 279 C 280 B 281 C
D. In-vitro IDO2 Enzyme (Indoleamine 2,3-dioxygenase2) Assay
[0781] Human indoleamine 2,3-dioxygenase-2 (hIDO2) catalyzes the
oxidative cleavage of the pyrrole ring of the indole nucleus of
tryptophan to yield N-formylkynurenine which can be converted to
kynurenine (KYN) by deformylation. hIDO2 with an C-terminal His tag
(Sino Biological Inc (China)) was expressed in E. coli cells and
the protein was purified using standard methods well known in the
art. Unless otherwise stated, all materials were procured from
Sigma Aldrich, Mo., USA.
[0782] The assay monitoring the conversion of L-tryptophan to KYN
was carried out as follows. hIDO.sub.2 (160 nM) was incubated with
tryptophan (5000 04) in the presence of ascorbic acid (20 mM),
methylene blue (10 04) and catalase (100 .mu.g/mL) in potassium
phosphate buffer (100 mM; pH 7.5) at 37 degrees C. for 30 min. The
reaction was terminated with 30% trichloroacetic acid (TCA) and
further incubated at 65 degrees C. for 15 min to fully convert
N-formylkynurenine to KYN. The reaction mixture was then
centrifuged to remove sediments, and the KYN in the supernatant was
measured by UV-visible absorption spectroscopy at 360 nm using a
Waters HPLC system fitted with a C-18 column or by LC/MS/MS (C. J.
D Austin et. Al, Amino Acid 2009, 565-578).
[0783] Percent inhibition at each concentration of test compounds
was determined by determining the decrease in KYN with reference to
the reaction control with 1% DMSO vehicle. Data were analyzed using
nonlinear regression to generate IC50 values using Graph Pad
Prism.RTM. 5. Compounds 2 and 184 were tested as described above.
Compounds 2 and 184 inhibit IDO2 activity with an IC50 less than 1
uM and reduced the levels of kynurenine pathway metabolite KYN
(Table 5).
TABLE-US-00008 TABLE 5A IC50 < 1000 nM; B: IC50 = 1000 to 2000
nM; C: IC50 > 2000 nM IDO2 Enzyme assay IC50 values No. IDO2 2 A
184 A
[0784] As described herein in example 30 and TABLES 3-5, the
compounds of the invention inhibit one or more of IDO1 or IDO2 or
TDO.
Example 31
Reduction of LPS Induced Plasma Kynurenine Levels in C57BL/6
Mice
[0785] Inflammatory mediators such as Lipopolysaccharides (LPS) and
Interferon-gamma (TFNg) arc well-established inducers of IDO1
expression. Intraperitoneal (i.p.) administration of bacterial
lipopolysaccharide (LPS) induces peak IDO1 activity in a variety of
tissues within one day after LPS administration resulting in the
production and release of kynurenine into the bloodstream
(Takikawa, O., et al. (1986) J. Biol. Chem. 261:3648-53; Yoshida,
H., et al. (1998) Cell 94:739-750). LPS-injected mice have been
used as models to study IDO1 expression and activity. Three eight
fed C57 BL/6 mice (age 7-8 weeks, weight: about 20-22 g) were
injected intrapritoneally with bacterial lipopolysaccharide (LPS;
26:B6 Sigma) at a concentration of 6 mg/kg. Animals were then
housed in normal condition for 20 hours at which time the test
compounds were administered orally in formulation containing 30%
polyethylene glycol 400 (PEG 400) and 20% propylene glycol (PG) in
normal saline (Dosing volume 10mL/kg). Blood was drawn through
retrorbital bleeds into a tube containing 100 mM EDTA for plasma
collection at the following times: just prior to LPS treatment,
just prior to test compound dosing (0 hr) and then at 2 hr, 4 hr, 6
hr, 8 hr, 24 hr and 48 hr post-test compound dosing. Plasma KYN and
drug levels were determined by LC/MS/MS using an API4000 mass
spectrometer (Applied Biosystems) coupled to a Shimadzu Prominence
LC system fitted with a C18 column.
[0786] Compounds were tested as described above and the data is
shown in TABLE 6. In vivo pharmacodynamic studies with LPS-injected
mouse model show that the compounds of the invention inhibit the
activity of IDO1 and reduce plasma kynurenine metabolite, KYN
levels in vivo.
TABLE-US-00009 TABLE 6 A > 50%, B > 25% < 50%, C < 25%
No. activity Formulation in NS 2 A 30% PEG & 30% PG 26 A 25%
DACM & 37.5% PG 58 A 30% PEG & 20% PG 59 A 40% PEG, 20% PG
& 10% DACM 92 A 30% PEG & 20% PG 95 B 30% PEG & 30% PG
96 A 30% PEG & 20% PG 97 A 30% PEG & 20% PG 101 A 30% PEG
& 20% PG 103 A 30% PEG & 30% PG 117 B 30% PEG & 20% PG
121 C 40% PEG & 20% PG 140 A 30% PEG & 30% PG 142 A 30% PEG
& 30% PG 147 A 10% CrEL, 10% EtOH & 20% PG 149 A 10% CrEL,
10% EtOH & 20% PG 154 A 15% CrEL, & 15% EtOH 158 A 30% PEG
& 20% PG 164 A 30% PEG & 20% PG 166 A 30% PEG & 20% PG
169 A 10% CrEL, 10% EtOH & 20% PG 170 B 40% PEG, 20% PG &
10% DACM 171 A 40% PEG, 20% PG & 10% DACM 171 A 40% PEG, 20% PG
& 10% DACM 177 A 40% PEG, 20% PG & 10% DACM 180 A 40% PEG,
20% PG & 10% DACM 181 A 40% PEG, 20% PG & 10% DACM 183 A
15% CrEL & 15% EtOH 184 A 30% PEG & 20% PG 188 A 30% PEG
& 20% PG 190 B 30% PEG & 20% PG 192 A 40% PEG & 20% PG
198 A 40% PEG & 20% PG 204 B 40% PEG & 20% PG 207 A 40% PEG
& 20% PG 208 A 40% PEG & 20% PG 213 B 40% PEG & 20% PG
234 B 30% PEG & 20% PG 240 A 30% PEG & 20% PG
Example 32
A. In Vivo Testing of KYN Pathway Inhibitors for Antitumor
Activity
[0787] Tumor volume reduction by the test compounds was evaluated
in a syngeneic CT26/Balb/C tumor model as described below. Balb/c
mice were purchased from Vivo Bio Tech, Hyderabad, India, and
housed in sterile conditions using an individually ventilated
caging system manufactured by Citizen Industries Limited,
Ahmedabad, India. Mice were quarantined for at least 7 days before
experimentation.
[0788] Mouse colorectal tumor forming cells, CT-26 cells (ATCC,
USA), were cultured in DMEM and 10% FBS medium supplemented with lx
non-essential amino acid (HiMedia) in an incubator maintained at
37QC with 95% humidity and 5% CO.sub.2. One million
(1.times.10.sup.6) live syngeneic CT-26 cells suspended in 0.2 ml
of 1.times. FBS were injected subcutaneously in the right flank of
each Balb/c mouse (age 6-8 weeks, weight: about 18-20 g) on day 1
under anesthesia. Eight to ten mice were used per experimental
group. Subcutaneous cells formed localized tumors. Animals were
visually inspected twice a day. Tumor measurements were initiated 7
days post injection using Vernier calipers. Tumor sizes at day 7
ranged typically from 80-150 mm.sup.3. Tumor dimensions were
subsequently measured every 3 to 4 days and tumor volume was
determined. Assuming tumors to be ellipsoid, tumor volume was
calculated using the formula:
V=(D.times.d.times.d)/2
[0789] Where D is longest diameter in mm, and d is shortest
diameter in mm.
[0790] When the average tumor volume in the mice reached 100-120
mm.sup.3, tumor bearing mice were randomized for treatment with
either test compound or vehicle alone. Test compounds were
formulated generally in normal saline comprising Polyethylene
Glycol (PEG400), Propylene glycol (PG) and Dimethyl Acetamide
(DACM) at pH 4.5-5.0 and dosed orally at ranges 40-75 mg/kg (dose
volume-5 mL/kg) twice a day. Solutions of the compounds were
prepared immediately before dosing the animals. In general, tumor
volume was measured at various times post-dosing from day 1 to day
34.
[0791] A non-limiting example is provided. Tumor bearing mice were
dosed orally twice a day at 40 mg/kg body weight with either
vehicle control or test compound 2 formulated in 30% Polyethylene
Glycol (PEG-400), 20% Propylene glycol (PG) in normal saline.
Tumors were measured and tumor volumes were determined at 1, 4, 6,
9, 11, and 14 days after the first dose as described above. Animals
were sacrificed on day 14. Tumor and blood samples were harvested
for further analysis. Just prior to animal sacrifice, blood was
drawn retro-orbitally into a tube containing 0.2% EDTA for plasma
collection. The harvested tumors were weighed and snap frozen in
liquid nitrogen immediately. Plasma KYN concentrations are measured
for pharmacokinetic analysis using LC/MS/MS using an API4000 mass
spectrometer (Applied Biosystems) coupled to a Shimadzu Prominence
LC system fitted with a C18 column.
[0792] Compound 2 significantly decreased tumor growth in compound
2-treated animal over 14 days compared to control vehicle-treated
animals (FIG. 1(a)). Similarly, mice dosed with compounds 97, 166
and 184 reduced tumor growth by 26%-38% (FIG. 1(b)-1(d)) compared
to control animals. Plasma KYN levels were reduced by 40-50% in the
compound treated animals compared to control animals (data not
shown).
B. Combination Therapy with Chemotherapy
[0793] Test compounds were evaluated in mice syngeneic tumor models
described above for usefulness in combination therapy with a
chemotheraputic agent, Doxorubicin (DOXO, Sterling Biotech,
India).
[0794] Tumor bearing Balb/c mice as described earlier were
randomized to receive vehicle alone, test compound alone, DOXO
alone, or test compound in combination with DOXO. Animals in DOXO
treated arm were injected intraperitoneally on day 1 with DOXO
formulated in normal saline (7.5 mg/kg). Five days later, the
animals were dosed twice a day orally with either a test compound
or vehicle alone during the test period till the end of the
experiment. A second smaller dose of DOXO at 3 or 4 mg/Kg was
administered to the DOXO-treated animals about 3 weeks after the
first DOXO dose. Tumor volume was measured in animals as described
above.
[0795] Non-limiting examples follow. Test compound 97 was
formulated in 40% PEG, 20% PG, 10% DACM in normal saline. From day
5 after the first DOXO dose was administered till the end of the
study period on day 35, the animals were dosed twice a day orally
with compound 97 (75 mg/kg). Three weeks later, the DOXO-treated
animals were injected i.p. with a second dose of DOXO (3 mg/kg).
Tumor volume was monitored as described above. Tumor tissues and
final blood samples were harvested from the mice 4 hours after the
final test compound dose. Tumor samples were weighed and snap
frozen as described previously. Animals treated with combination of
compound 97 and DOXO showed greater reduction in tumor volume than
that achieved by DOXO or by test compound alone (FIG. 2(a)).
[0796] Similarly, a second set of experiments was conducted with
test compound 184 formulated in 40% PEG, 20% PG, 10% DACM in normal
saline. As described above, animals were dosed with test compound
184 at 50 mg/kg twice a day orally during the study period. Animals
treated with a combination of compound 184 and DOXO showed greater
reduction in tumor volume than that achieved by DOXO or by the test
compound alone (FIG. 2(b)). Thus, the time of tumor progression is
reduced by treatment with compounds of the invention.
C. Tissue Penetration Assays
[0797] i. Tumor penetration in mice: Tumor penetration of test
compounds and KYN levels in tumor and plasma of treated animals
were determined in a syngeneic CT26/Balb/C tumor model. Briefly,
three to four tumor bearing Balb/c mice (tumor size=150-200
mm.sup.3) were dosed twice a day orally for 4 days with test
compound (for e.g., compound 2) at 50 mg/kg formulated in
carboxymethyl cellulose (CMC). A vehicle control was included along
with the test compounds. Blood samples were collected by
retro-orbital bleed or terminal cardiac bleed into tubes containing
heparin just before the final dose and at two hours after the final
dose. Animals were sacrificed two hours post-final dose. Tumors
were harvested and weighed immediately. Tumors were then snap
frozen in liquid nitrogen, pulverized and dissolved in
homogenization buffer (chilled acetonitrile/water/formic acid
(10:90:0.1 v/v/v) to extract test compounds and kynurenine,
KYN.
[0798] Plasma and tumor tissue KYN levels were reduced in test
compound treated animals compared to control vehicle-treated
animals (TABLE 7). For example, compounds 2 and 97 each reduced
tumor KYN levels by 81%. At 2 hours post-dosing, the plasma KYN
levels were reduced by 62% and 67% respectively in the animals
treated with compound 2 or 97 compared to the control animals dosed
with vehicle alone. The concentration of test compounds in the
tumor tissue extracts is determined by LC/MS/MS using an API 4000
or AP15500 Qtrap mass spectrometer.
TABLE-US-00010 TABLE 7 Dose % KYN drop No (mg/kg) Plasma Tumor 2 50
A A 142 50 A A 26 20 B A 97 50 A A 98 50 C C 103 50 B C 145 50 B C
154 50 A B 158 50 A A 164 50 B B 166 50 B B 181 50 B C 183 50 B C
180 50 C B 184 50 A A 188 50 B A 190 50 B A 195 50 A A 198 50 A A
204 50 A A
[0799] ii. Brain penetration in rats: Early and accurate prediction
of drug penetration of the blood brain barrier is vital for the
development of drugs targeting the central nervous system.
Accordingly, ability of the compounds of the invention to penetrate
the blood brain barrier was tested according to the methods well
established in the art (Giorgetti et al., (2010) JPET 333:748-757,
2010). Briefly, Sprague-Dawley rats (n=3 per time point studied)
were administered a single intravenous dose of the test compounds 2
or 97 formulated in 40% PEG and 20% PG at 10 mg/kg. Three rats were
used for each of the following time points post-dosing: 0.5 hr, 1.5
hr and 3.0 hrs. First, blood was collected during deep anesthesia
in heparinized tubes, and then animals were sacrificed and their
brains were harvested. The brains were then pulverized in liquid
nitrogen and homogenized in fixed volumes of ice cold
homogenization buffer (10% Acetonitrile, 0.1% formic acid in
water). Plasma was separated from blood using standard procedures.
Plasma and brain concentrations of test compounds 2 and 97 were
determined by LC/MS/MS using an API 4000 or AP15500 Qtrap mass
spectrometer. Brain/Plasma ratio of test compounds 2 and 97 were
determined after normalizing brain weight and buffer volume.
[0800] Compound 2 and 97 are capable of penetrating blood brain
barrier to enter the brain tissue (TABLE 8). Accordingly, the
compounds of the invention are useful for treating kynurenine
pathway- and/or IDO1- and/or IDO2- and/or TDO-associated brain
diseases.
TABLE-US-00011 TABLE 8 Ratio of Drug Concentration No. Time (hr)
(Brain:Plasma) 2 0.5 9.8:1 1.5 10.7:1 3.0 6.0:1 97 0.5 0.67:1 1.5
0.47:1 3.0 0.64:1
Example 33
Induction of T-Cell Proliferation
[0801] Conceptually, a reduction in the immune suppressive effects
of kynurenine pathway metabolites or one or more of IDO1 or IDO2 or
TDO enzymes may result in increased numbers or reactivity of tumor
specific immune cells. Further, inhibition of one or more of IDO1
or IDO2 or TDO enzymes may further increase the number or
reactivity of tumor reactive immune cells when combined with other
therapeutics, for example chemotherapeutics (e.g., doxorubicin)
and/or immune modulators (e.g., anti-CTLA4 and/or anti-PD-L1 and
/or anti-PD1 antibody). In animal models described below, it may
also be possible to directly and/or indirectly measure the number
and/or activity of tumor reactive immune cells. Methods for
measuring the number and/or activity of tumor reactive immune cells
are well established and can be performed using techniques familiar
to those skilled in the art (Current Protocols in Immunology, vol
4, Coligan, J. E., et al; Immunotherapy of Cancer, Human Press,
2006, Disis, M. L. and references therein). The following
experiment addresses the ability of the compounds of the invention
to induce T-cell proliferation and reduce the immunosuppressive
effects of kynurenine pathway metabolites or the activity o one or
more of IDO1 or IDO2 or TDO enzymes.
[0802] Effect of Kynurenine Pathway Inhibitors on T-Cell
Proliferation that is Suppressed by IDO1-Expressing Dendritic
Cells
[0803] Monocytes are collected from human peripheral blood
mononuclear cells by leukophoresis. Monocytes are then seeded at a
density of 1.times.10.sup.6 cells/well in a 96 well plate, using
RPMI 1640 medium supplemented with 10% FBS and 2 mM L-glutamine
(all from Invitrogen). Adherent cells arc retained on the plate
after overnight culture at 37.degree. C. Adherent monocytes are
then stimulated for 5-7 days with 100 ng/ml GM-CSF (PeproTech) and
(250 ng/ml L4 (PeproTech), followed by activation with 0.5 ug/ml
LPS from Salmonella typhimurium (Sigma) and 50 ng/mL IFN gamma
(R&D systems) for additional 2 days to induce dendritic cell
maturation.
[0804] After dendritic cell activation, the medium is replaced with
completed RPMI 1640 supplemented with 100-200 U/ml L-2
(ProSpec-Tany Technogene) and 100 ng/mL anti-CD.sub.3 antibody
(PharMingen), T-cells (-3 times.10.sup.5 cells/well), and serial
dilutions of test compounds. After incubation for 2 more days,
T-cell proliferation are measured by BrdU incorporation assay,
using a colorimetric Cell Proliferation ELISA kit per
manufacturer's instruction (Roche Molecular Biochemicals). Cells
are continuously cultured for 16-18 hours in the presence of 10 uM
BrdU labeling solution. The FixDenat solution is removed and 100
ul/well ant-BrdU-POD antibody conjugate working solution was added.
The reaction is carried out at room temperature. The antibody
conjugate is then removed and cells were rinsed 3 times with 200
ul/well washing solution. Finally, 100 ul/well of substrate
solution is added and the results are obtained using a microreader
(Spectra Max Plus, Molecular Devices) during color development.
Multiple readings at various time points are obtained to ensure
that the data is within the linear range. The data is routinely
obtained from replicated experiment, and appropriate controls are
included. (Terness P, et al. (2002) J. Exp. Med. 196(4):447-57; and
Hwu P, et al. (2000) J. Immunol. 164(7):3596-9).
Example 34
[0805] In Vivo TDO Assay
[0806] The ability of test compounds to inhibit TDO activity in
vivo in tumors can be determined using P815 mouse tumor model
(Uyttenhove C, et al. (2003) Nat Med 9:1269-1274; Pilotte L, et al.
PNAS 2012). Briefly, P815 tumor cells are transfected with TDO
cDNA. TDO-expressing tumor cells arc injected i.p. into naive
syngenic DBA/2 mice (Harlan, USA) resulting in tumor growth. Tumor
volumes arc measured as described above. When the average tumor
volumes in the mice reach 100-120 mm.sup.3, tumor bearing mice are
randomized for treatment with either test compound or vehicle
alone. Test compounds are formulated generally in normal saline
comprising Polyethylene Glycol (PEG400), Propylene Glycol (PG) and
Dimethyl Acetamide (DACM) at pH 4.5-5.0 and dosed orally at ranges
40-75 mg/kg (dose volume-5 mL/kg) twice a day. Solutions of the
compounds are prepared immediately before dosing the animals. In
general, tumor volume is measured at various times post-dosing from
day 1 to day 34. The reduction in tumor progression and plasma Kyn
levels are determined as described above.
[0807] Liver and tumor tissue homogenates are obtained from the
mice treated with either a test compound or vehicle. Assays for
hepatic and tumor TDO activity are carried out using L-Trp as
substrate as previously described (Pilotte L, et al. PNAS 2012),
with activity expressed as .mu.mol of KYN formed per hour per gram
of wet liver weight. Briefly, dry pellets of liver and tumor
tissues are lysed in 50 mM potassium phosphate buffer (pH 7.5). 150
mM KCL, 250 mM sucrose, 1 mM L-Tryptophan, 10 uM bovine hemin and
protease inhibitors (complete EDTA free, Roche Applied Science).
The extract is first centrifuged at 4 degree C. for 5 min at
700.times.g and the supernatant is centrifuged at 20,000.times.g
for 15 min. The buffer of the clarified extract is then exchanged
over 50 mM potassium phosphate buffer (H 7.5) using a HiTrap
desalting column (GE Healthcare), and aliquots are frozen in liquid
nitrogen and kept at -80 C until further use.
[0808] TDO activity is measured as follows: The reaction mixture
contain (final concentration) 50 mM potassium phosphate buffer (pH
7.5), protease inhibitors, 20 mM ascorbic acid, 10 uM methylene
blue, 500 units/ml catalase (bovine liver, Sigma), and L-Tryptophan
in the presence or absence of test compounds. The reaction is
initiated by addition of 100 .mu.l of cell/tissue extract to 100
.mu.l of reaction mixture prewarmed at 37 C. The reaction is
conducted at 37 C for 10, 30, 60 min and stopped by the addition of
40 .mu.l of TCA 30% (wt/vol). To convert N-formylkynurenine to
kynurenine, the reaction mixture is mixed with 125 .mu.l of 2%
(wt/vol) 4-(Dimethylamino) benzaldehyde in acetic acid and
incubated for 10 min at room temperature. The reaction mixture is
then centrifuged to remove sediments and the supernatant was
monitored by LC/MS/MS using a API4000 mass spectrometer (Applied
Biosystems) coupled to a Shimadzu Prominence LC system fitted with
a Cl 8 column. Percent inhibition at each concentration of test
compounds is determined by estimating the decrease in KYN. Data are
analyzed using nonlinear regression to generate IC.sub.50 values
using Graph Pad Prism.RTM. 5.
Example 35
In Vivo Testing of Inhibitors in LPS-Induced Depressive-Like
Behaviors in Mice Model
[0809] Increased levels of Kynurenine pathway metabolites,
independent of immune activation or IDO1 activation, are capable of
inducing depressive-like behaviors in mice administered with
increasing doses of exogenous L-Kynurenine. LPS administration is
known to activate IDO1 and culminate in depressive-like behavioral
syndrome. Depressive behavior of mice in LPS injected mice has been
characterized by increased duration of immobility in both the
forced swim and tail suspension tests. Compounds of the invention
reduce plasma KYN levels (Examples 31 and 32; Tables 6 and 7). The
ability of the compounds of the invention to reduce depressive-like
behavior are tested in mice injected with LPS that show increased
KYN levels and increased IDO1 and/or TDO activity following
procedures well known in the art (O'Connor et al. Mol. Psychiatry.
2009 May; 14(5): 511-522; Porsolt, R. D. Rev Neurosci.
2000;11(1):53-8).
[0810] Behavioral Experiments--Locomotor Activity
[0811] The effects of LPS on locomotor activity (LMA) is assessed
in mice individually placed in a clean, novel cage similar to the
home cage, but devoid of bedding or litter. The cage is divided
into four virtual quandrants, and the LMA is measured by counting
the number of line crossings and rearings over a 5 min period.
Counting is done by a well-trained observer who is blinded to the
treatments.
[0812] Behavioral experiments--Forced Swim Test
[0813] The forced swim test (FST) is conducted according to methods
well established and known in the art (Porsolt, R. D. Rev Neurosci.
2000; 11(1):53-8). Briefly, each mouse is placed individual in a
cylinder (diameter 23 cm; height 31 cm) containing 15 cm of water
maintained at 23+/-1 degree C. The water is changed between testing
sessions. Mice are placed into the water for 6 min and then
returned to their home cage. During the test, the mice are video
recorded from above, and the duration of immobility is determined
over the last 5 min of the test using the mobility function of the
"Observer Basic" software (Nolus, Netherlands). Briefly, the mice
arc recognized in contrast from their background and tracked in 2
different dimensions as the surface arc of the detectable object
(mouse) moves within the predefined arena. Mobility is defined as
the displacement of detectable surface area (mouse) over time and
is averaged over 3 sample intervals to reduce error generated by
sharp movements or missing frames in the digital record. Program
analysis settings can be: Sampling rate=3/s; detection
method=substration with low threshold of 20 and high threshold of
255 and minimum detectable object size of 200 pixels; image
filtering=2 pixel erosion and dilation; mobility threshold of 20%
with 3 interval averaging. It is well understood that other
settings can be used by those of skill in the art.
[0814] Behavioral Experiments--Tail Suspension Test
[0815] The mice are taken from their home cage and a small piece of
adhesive tape is placed approximately 2 cm from the tip of the
tail. A single hole is punched in the tape and the mice are hung
individually for a period of 10 min on a hook connected to a strain
gauge. A computerized system for processing the force exerted on
the guage (Mouse Tail Suspension Package, MED-TSS-MS, Med
Associates, St. Albans, Vt.) automatcally collected and analyzed
the movements of each individual mouse. The time of immobility is
determined after establishing a threshold of each individual mouse
that is set precisely at the activity level that would exclude all
movements and only encompass immobility. Time below the threshold
indicates the time of immobility. Program analysis settings can be:
integration=on; resolution=0.1; gain=4; start rigger=20.
[0816] Behavioral data are analyzed using a one-way (treatment),
two-way (pretreatment X treatment) or a three-way (pretreatment X
treatment X time) ANOVA with repeated measurement on the time
factor where appropriate, followed by a post-hoc pairwise multiple
comparison procedure using the Fisher's LSD method, if the
interaction is significant.
EXAMPLE 36
[0817] In Vivo Testing of the Inhibitors in Human Immunodeficiency
Virus-1 (HIV-1) Encephalitis Model
[0818] Inhbition of IDO1 is known to enhance the elimination of
HIV-1 virus infected macrophages in the central nervous system in
animal models of a Human Immunodefficency Virus 1 (HIV-1)
Encephalities (Potula et al. Blood. (2005) 106: 2382-2390). The
ability of the test compounds to eliminate infected HIV-1
macrophages effectively in the nervous system can be tested using
such well-established models.
Cell Isolation and Viral Infection
[0819] Monocytes and PBL can be obtained by counter current
centrifugal elutriation of leukopheresis packs from Dulbecco's
Modified Eagle's Medium (DMEM, Sigma-Aldrich) supplemented with 10%
heat-inactivated pooled human serum, 1% glutamine, 50 ug/ml
gentamycin, l0ug/m1 Ciproflaxin (Sigma), and 1000 U/ml highly
purified recombinant human macrophage colony-stimulating factor.
After 7 days in culture, MDM are infected with HIV-lADA at
multiplicity of infection of 0.01.
Hu-PBL-NOD/SCID HIVE Mice 4 week old male NOD/C.B-17 SCID mice can
be purchased from a resource such as Jackson Laboratory (Bar
Harbor, Minn., USA). Animals are maintained in sterile
microisolator cages under pathogen-free conditions. All animals are
injected i.p. with rat anti-CD.sub.122 (0.25 mg/mouse) 3 days
before PBL transplantation ad two with rabbit asialo-GM1 antibodies
(0.2 mg/mouse) (Wako) one day before and three days after PBL
injection (20.times.10.sup.6 cells/mouse). HIV-1ADA infected MDM
(3.times.10.sup.5 cells in 10 ul) are injected intracranially
(i.c.) eight days following PBL reconstitution generating
hu-PBL-NOD/SCID HIVE mice. Immediately following the intracranial
injection of HIV-1 infected MDM the hu-NOD/SCID mice are
subcutaneously (s.c.) implanted with control (vehicle) or compound
pellets (14-28 day slow release, Innovative research). Experiments
are desiged to confirm the induction of virus-specific CTLs in
hu-NOD/SCID mice by tetramer staining and neuropathic analyses of
MDM elimination from the brain tissue. Then, experiments are
designed to analyze human lymphocyte reconstitution, humoral
responses, and neuropathological alterations. In these experiments,
animals are bled on day 7 and sacrificed at 14 and 21 days after
i.c. injection of human MDM. Blood collected in EDTA-containing
tubes is used for flow-cytometry and plasma is used for detection
of HIV-1 p24 using ELISA (Beckman Coulter). HIV-1 specific
antibodies are detected by Western blot tests according to the
manufacturer's instructions (Cambridge Biotech HIV-1 Western Blot
kit, Calypte Biomedical). Similar amount of virus-specific
antibodies are detected in control and compound-treated animals. A
total of 3 independent experiments can be performed using 3
different human leukocyte donors.
FACScan of Peripheral Blood and Spleen in hu PBL-NOD/SCID HIVE
Mice
[0820] Two-color FACS analysis can be performed on peripheral blood
at weeks 1-3 and splenocytes at weeks 2 and 3 after i.c. injection
of human MDM. Cells are incubated with fluorochrome-conjugated
monoclonal antibodies (mAbs) to CD4, CD, CD56, CD3, IFN-gamma
(Bioscience) for 30 mins at 4 degrees C. To evaluate the cellular
response, IFN-gamma intracellular staining is performed in
combination with anti-human CD.sub.8 and FITC-conjugated anti-mouse
CD.sub.45 to exclude murine cells. To determine the
antigen-specific CTL, allophycocyainn-conjugated tetramer staining
for HIV-1.sub.gag (p17(aa77-85) SLYNTVATL, SL-9) and HIV-1.sub.pol
[(a476-485) ILKEPVHGV, L-9] is performed on phytohemaglutinin/IL-2
(PHA/IL2)-stimulated splenocytes. Cells are stained following the
recommendation of the NIH/NIAID, National Tetramer Core Facilities.
Data are analyzed with a FACS Calibor using CellQuest Software
(Becton Dickinson Immunocytometry System).
Histopathology and Image Analyses
[0821] Brain tissues are collected at days 14 and 21 after i.c.
injection of MDM, fixed in 4% phosphate-buffered paraformaldehyde
and embedded I paraffin or frozen at -80 degrees C. for later use.
Coronal sections from the embedded blocks are cut in order to
identify the injection site. For each mouse, 30-100 (5 um thick)
serial sections are cut from the human MDM injection site and 3-7
slides (10 sections apart) are analyzed. Brain sections are
deparaffinized with xylene ad hydrated in gradient alcohols.
Immunohistochemical staining follows a basic indirect protocol,
using antigen retrieval by heating to 95 degree C. in 0.01 mol/L
citrate buffer for 30 mi for antigen retrieval. To identify human
cells in mouse brains, mAb to Vimentin (1:50, clone 3B4, Dako
Corp.), which identifies all human leukocytes is used. Human MDM ad
CD.sub.8+lymphocytes are detected with CD.sub.68 (1:50 dilution,
clone KP 1) and CD.sub.8 (1:50 dilution, clone 144B) antibodies
respectively. Virus-infected cells are labeled with mAb to
HIV-1.sub.p24 (1:10, clone Kal-1, all from DAKO). Reactive murine
microglial cells are detected with Iba-1 antibody (1:500, Wako).
Expression of human IDO1 (huIDOl) and/or human TDO (huTDO) are
visualized with Abs obtained from Enzo Lifesciences (USA) and Abcam
(USA). Primary antibodies are detected with appropriate
biotinylated secondary antibodies and visualized with aviden-biotin
complexes (Vectastain Elite ABC kit, Vector Laboratories) or
horseradish-peroxidase (HRP)-coupled dextran polymer (EnVision,
DAKO). Immunostained sections are counterstained with Mayer's
hemotoxylin. Sections from which primary antibody is deleted or
irrelevant IgG isotype is incorporated serves as controls. Two
independent observers in a blinded fashion count the numbers of
CD.sup.8+ lymphocytes, CD.sup.68+ MDM, and HV-1.sub.p24 cells in
each section form each mouse. Light microscopic examination is
performed with a Nikon Eclipse 800 microscope (Nikon Instruments
Inc.). Semi-quantitative analysis for Ibal (percentage of area
occupied by immunostaining) is carried out by computer-assisted
image analysis (Image-Pro..RTM..Plus, Media Cybernetics).
[0822] Data is analyzed using Prism (Graph Pad) with Student t-test
for comparisons and ANOVA. P values<0.5 arc considered
significant. (Potula et al. Blood Oct. 1, 2005, vol: 106(7)
2382-2390).
Example 37
[0823] Increased levels of IDO1 and kynurenine pathway metabolites
has been observed in the autopsy brain slices brains of Alzheimer's
disease subjects (Bonda et al. Redox Rep. 2010; 15(4): 161-168) and
circulating antibodies to the kynurenine metabolites have also been
identified (Duleu et al. International Journal of Alzheimer's
Disease, vol. 2010, Article ID 501541, 6 pages, 2010.
doi:10.4061/2010/501541). The ability of the test compounds to
reduce the kynurenine pathway metabolites in the brains by
inhibiting kynurenine pathway or one or more of IDO1 or IDO2 or TDO
is tested used animal models of Alzheimer's disease.
[0824] Various Alzheimer's disease mouse models animals (for
example, B6; 129-Psen1tm1Mpm Tg(APPSwe,tauP301L)1Lfa/Mmjax,
B6C3-Tg(APPswe, PSEN1dE9).sub.85Dbo/Mmjax ,
B6.Cg-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax, B6.129-Tg(APPSw)40Btla/Mmjax,
B6.Cg-Nos2tm1Lau Tg(Thy1APPSwDutIowa)BWevn/Mmjax) can be purchased
from Jackson Laboratories, USA. As an non-limiting example, B6;
129-Psen1tm1Mpm Tg(APPSwe, tauP301L)1Lfa/Mmjax mice are purchased
and housed in sterile conditions as described before. The mice
develop neurofibrillary tangles in the hippocampus after about 6
months and in the rest of the cortex subsequently. Cognitive
deficits in the mice are manifested about 4 months of age although
no neuronal loss has been reported in these mice.
[0825] Mice about 6 moths of age are randomized to be dosed in a
single intravenous dose of either a vehicle control or a test
compound formulated in PEG and PG in normal saline at 10 mg/kg as
described in earlier examples. In one set of experiments, the
ability of the compounds to penetrate blood brain barrier and
reduce plasma kynurenine metabolites by the test compounds tested.
Three mice are used for each of the following time points
post-dosing: 0.5 hr, 1.5 hr and 3.0 hrs. Brain and plasma drug and
KYN levels are determined in the animals as described previously.
Presence and reduction of circulating antibodies to kynurenine
pathway metabolites is determined (Duleu et al. International
Journal of Alzheimer's Disease, vol. 2010, Article ID 501541, 6
pages, 2010. doi:10.4061/2010/501541).
[0826] In a second set of experiments, effect of test compounds on
the cognitive function of the mice is tested. Test compound and
vehicle control treated animals are dosed orally twice as day as
previously described for 8 days and subjected to behavioral tests
that arc well known in the art (Choi et al. J. Neurochem. 2013,
124(1) 59-68).
[0827] Briefly, spatial leaming and memory is assessed using the
Morris Water Maze, as previously described (McKee et al. Brain Res.
2008, 1207, 225-236) in mice treated with either a test compound or
a vehicle control. Briefly, a circular plastic pool is filled with
water (22.degree. C.) that is colored with non-toxic white paint to
obscure the location of a submerged platform. Three visual cues are
placed around the tank to orient the mice, with the platform
remaining in a fixed location. The platform location is kept
constant for each mouse during training, and it is 1.5 cm beneath
the surface of the water. On each day, training consisted of five
trials. For each trial, the mouse is placed into the pool facing
the wall from one of four randomly varied start positions and
allowed to swim until finding the platform. If a mouse fails to
find the platform within 60 s, the mouse is manually guided to the
hidden platform and allowed to stay on the platform for 30 s. Probe
trials for retention of spatial training are conducted 1.5 and 24 h
after the last training trial. During the probe trials, the
platform is removed and mice are free to swim in the pool for 60 s.
Mice are monitored with a camera mounted on the ceiling directly
over the pool and recorded for subsequent analysis. The escape
latency to cross the platform location, the number of platform
location crosses, and path length are recorded. The ability of the
test compound to improve cognitive function and reduce plasma and
brain kynurenine metabolite levels is determined.
Example 38
Identification of Metabolites of the Test Compounds
[0828] Prodrugs and metabolites of a compound may be identified
using routine techniques known in the art. See, e.g., Bertolini et
al., J. Med. Chem., 40, 2011-2016 (1997); Shan, et al., J. Pharm.
Sci., 86 (7), 765-767; Bagshawe, Drug Dev. Res., 34, 220-230
(1995); Bodor, Advances in Drug Res., 13, 224-331 (1984);
Bundgaard, Design of Prodrugs (Elsevier Press 1985); Larsen, Design
and Application of Prodrugs, Drug Design and Development
(Krogsgaard-Larsen et al., eds., Harwood Academic Publishers,
1991); Dear et al., J. Chromatogr. B, 748, 281-293 (2000); Spraul
et al., J. Pharmaceutical & Biomedical Analysis, 10, 601-605
(1992); and Prox et al., Xenobiol., 3, 103-112 (1992).
[0829] Metabolites of test compounds are identified by in vitro
incubations with liver microsomes, liver S9 fractions and
hepatocytes using either cold or .sup.14C-labeled compounds. They
arc identified using HPLC based on retention times, MS and MS/MS
fragmentation times and NMR analysis.
A. Liver Microsomes
[0830] Test compounds are incubated with pooled liver microsomes
from mouse. Incubations are carried out at 37 degrees C. in a
shaking water bath. The incubation mixtures consist of: 0.1 M
Potassium Phosphate buffer, pH 7.4, 0.5 mM NADPH, 0.5 mg/ml liver
microsomal protein, and 20 uM of a test compound. After 5 mins
pre-incubation at 37 degree C., to incubations are initiated with
the addition of the test compound and NADPH. After 60 mins, the
mixtures are quenched by adding one volume of cold acetonitrile.
The samples are vortexed and centrifuged for 10 mins at 14000 rpm.
Aliquots of the supernatant (200 ul) are taken for LC/MS/MS
analysis.
B. Liver S9 Fractions
[0831] The test compounds are incubated at 37 degree C. with pooled
liver S9 fractions taken from humans in a shaking water bath. The
incubation mixtures consist of: 0.1M Potassium Phosphate buffer, pH
7.4, 0.5 mM NADPH, 0.5 mg/ml liver microsomal protein, and 20 uM of
a test compound. After 5 mins pre-incubation at 37 degree C., to
incubations are initiated with the addition of the test compound
and NADPH. After 2 hours, the mixtures are quenched by adding one
volume of cold acetonitrile. The samples are vortexed and
centrifuged for 10 mins at 14000 rpm. Aliquots of the supernatant
(200 ul) are taken for LC/MS/MS analysis.
C. Hepatocytes
[0832] A test compound is incubated with pooled hepatocytes from
mice at concentrations of 3 uM to 30 uM. The incubation mixtures
are prepared by adding 4.5 ul/ml of 0.67 and 6.7 uM of .sup.14
C-labeled compound stock solutions in acetonitrile for 3 uM to 30
uM incubations respectively at an appropriate amount of incubation
medium (2 to 5 ml) containing a 10.sup.6 cells/ml. All samples are
incubated at 37 degrees C. for 3 hours in an incubatr under 5%
CO.sub.2 atmosphere on an orbital shaker. The mixtures are quenched
by adding one volume of cold acetonitrile. The samples are vortexed
and centrifuged for 10 mins at 14000 rpm. Radioactivity in 20 ul of
supernatant is measured by liquid scintillation and 20 ul of the
suprnatents is injected into HPLC for metabolite profiling.
[0833] Metabolites of test compounds can also also identified from
in vivo urine, bile, feces and plasma samples from mammals
including humans using .sup.14 C-labeled test compounds. A
representative study is described below.
[0834] Six mice arc dosed orally with a .sup.14 C-labeled test
compound with a target dose of 100 mg/kg, 100 uCi/kg. The dosing
solutions are prepared in 50 mM citrate buffer the night before
dosing and stored at RT in the dark. Bile, urine and feces are
obtained over intervals through 48 hours post-dose. Blood is drawn
before dosing and at 0.5, 1, 2, 4, 8, 12, 24 and 48 hours post
dosing. Plasma is prepared from blood samples by centrifuging for
15 min at 1000 g.
A. Bile
[0835] A representative pooled bile sample is prepared by combining
5% portion of volume of each bile sample/mouse collected over 48
hours. The pooled bile sample is diluted 1:1 (vol:vol) by addition
of water to bile. The diluted bile is centrifuged for 10 min at
14000. Supernatants (20 ul aliquot) are analyzed by LC/MS/MS for
the metabolite identification.
B. Urine
[0836] A representative pooled urine sample is prepard by combining
20% portion of volume of each urine sample/mouse collected over 48
hours. The pooled urine sample is diluted 1:1 (vol:vol) by addition
of water to bile. The diluted bile is centrifuged for 10 min at
14600. Supernatants (30 ul aliquot) are analyzed by LC/MS/MS for
the metabolite identification.
C. Plasma
[0837] Two pooled samples (1 h and 4 h) are prepared by combining
equal volumes of plasma samples from each mouse that are collected
for each time point. The pooled samples are each extracted once
with 5 ml acetonitrile/methanol (50:50 by volume) and twice
additionally with 3 ml acetonitrile/ethanol. After extraction with
organic solvents, the extracted samples are centrifuged at 4000 rpm
for 10 min at 5 degree C. and the supernatants from each
centrifugation step are combined. The supernatants are evaporated
to dryness at RT under N.sub.2 gas. The dried residues are
reconstituted in 350 ul of the mobile phase of 80%/20% water
containing acetonitrile. The samples are centrifuged again at 14000
rpm for 10 mins at 5 degrees C. 100 ul of supernatants are injected
into LC/MS/MS for metabolite identification.
D. Feces
[0838] A representative pooled feces sample is prepared by
combining 5% portion of volume of each feces sample/mouse collected
over 48 hours. The pooled feces sample (1 ml) is extracted with 2 m
of methanol/acetonitrile (50:50 vol/vol). The mixtures are
vortexed, sonicated for and centrifuged at 4000 rpm for 15 min.
Supernatants are collected and the pellets are extracted two more
times with methanol/acetonitrile/water (1:1:1 v/v/v) and the
supematents are combined and evaporated to dryness under N2. The
dried residues are suspended in 0.5 ml methanol/water 1:1 (v/v) and
centrifuged for 10 min at 14000 rpm. Supernatents (60 ul aliquot)
are analyzed by LC/MS/MS for the metabolite identification.
[0839] Without wishing to be bound to the above exemplar, it is to
be understood that in the methods described herein, the individual
components of a co-administration, combination can be administered
by any suitable means, contemporaneously, simultaneously,
sequentially, separately , alternation or in a single
pharmaceutical formulation. Where the co-administered compounds or
compositions are administered in separate dosage forms, dosage
levels, the number of dosages administered per day for each
compound may be the same or different. The compounds or
compositions may be administered via the same or different routes
of administration. The compounds or compositions may be
administered according to simultaneous or alternating regimens, at
the same or different times during the course of the therapy,
concurrently in divided or single forms.
[0840] All publications cited in this specification are
incorporated herein by reference. While the invention has been
described with reference to particular embodiments, it will be
appreciated that modifications can be made without departing from
the spirit of the invention.
[0841] Such modifications are intended to fall within the scope of
the appended claims.
* * * * *